Substrate Specificity: A characteristic feature of enzyme activity in relation to the kind of substrate on which the enzyme or catalytic molecule reacts.Molecular Sequence Data: Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories.Amino Acid Sequence: The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION.Kinetics: The rate dynamics in chemical or physical systems.Sensitivity and Specificity: Binary classification measures to assess test results. Sensitivity or recall rate is the proportion of true positives. Specificity is the probability of correctly determining the absence of a condition. (From Last, Dictionary of Epidemiology, 2d ed)Binding Sites: The parts of a macromolecule that directly participate in its specific combination with another molecule.Models, Molecular: Models used experimentally or theoretically to study molecular shape, electronic properties, or interactions; includes analogous molecules, computer-generated graphics, and mechanical structures.Catalysis: The facilitation of a chemical reaction by material (catalyst) that is not consumed by the reaction.Sequence Homology, Amino Acid: The degree of similarity between sequences of amino acids. This information is useful for the analyzing genetic relatedness of proteins and species.Catalytic Domain: The region of an enzyme that interacts with its substrate to cause the enzymatic reaction.Escherichia coli: A species of gram-negative, facultatively anaerobic, rod-shaped bacteria (GRAM-NEGATIVE FACULTATIVELY ANAEROBIC RODS) commonly found in the lower part of the intestine of warm-blooded animals. It is usually nonpathogenic, but some strains are known to produce DIARRHEA and pyogenic infections. Pathogenic strains (virotypes) are classified by their specific pathogenic mechanisms such as toxins (ENTEROTOXIGENIC ESCHERICHIA COLI), etc.Recombinant Proteins: Proteins prepared by recombinant DNA technology.Hydrolysis: The process of cleaving a chemical compound by the addition of a molecule of water.Base Sequence: The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence.Crystallography, X-Ray: The study of crystal structure using X-RAY DIFFRACTION techniques. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)Mutagenesis, Site-Directed: Genetically engineered MUTAGENESIS at a specific site in the DNA molecule that introduces a base substitution, or an insertion or deletion.Hydrogen-Ion Concentration: The normality of a solution with respect to HYDROGEN ions; H+. It is related to acidity measurements in most cases by pH = log 1/2[1/(H+)], where (H+) is the hydrogen ion concentration in gram equivalents per liter of solution. (McGraw-Hill Dictionary of Scientific and Technical Terms, 6th ed)Cloning, Molecular: The insertion of recombinant DNA molecules from prokaryotic and/or eukaryotic sources into a replicating vehicle, such as a plasmid or virus vector, and the introduction of the resultant hybrid molecules into recipient cells without altering the viability of those cells.Protein Binding: The process in which substances, either endogenous or exogenous, bind to proteins, peptides, enzymes, protein precursors, or allied compounds. Specific protein-binding measures are often used as assays in diagnostic assessments.Protein Structure, Tertiary: The level of protein structure in which combinations of secondary protein structures (alpha helices, beta sheets, loop regions, and motifs) pack together to form folded shapes called domains. Disulfide bridges between cysteines in two different parts of the polypeptide chain along with other interactions between the chains play a role in the formation and stabilization of tertiary structure. Small proteins usually consist of only one domain but larger proteins may contain a number of domains connected by segments of polypeptide chain which lack regular secondary structure.Sequence Alignment: The arrangement of two or more amino acid or base sequences from an organism or organisms in such a way as to align areas of the sequences sharing common properties. The degree of relatedness or homology between the sequences is predicted computationally or statistically based on weights assigned to the elements aligned between the sequences. This in turn can serve as a potential indicator of the genetic relatedness between the organisms.Protein Conformation: The characteristic 3-dimensional shape of a protein, including the secondary, supersecondary (motifs), tertiary (domains) and quaternary structure of the peptide chain. PROTEIN STRUCTURE, QUATERNARY describes the conformation assumed by multimeric proteins (aggregates of more than one polypeptide chain).Structure-Activity Relationship: The relationship between the chemical structure of a compound and its biological or pharmacological activity. Compounds are often classed together because they have structural characteristics in common including shape, size, stereochemical arrangement, and distribution of functional groups.Bacterial Proteins: Proteins found in any species of bacterium.Mutation: Any detectable and heritable change in the genetic material that causes a change in the GENOTYPE and which is transmitted to daughter cells and to succeeding generations.Species Specificity: The restriction of a characteristic behavior, anatomical structure or physical system, such as immune response; metabolic response, or gene or gene variant to the members of one species. It refers to that property which differentiates one species from another but it is also used for phylogenetic levels higher or lower than the species.Antibody Specificity: The property of antibodies which enables them to react with some ANTIGENIC DETERMINANTS and not with others. Specificity is dependent on chemical composition, physical forces, and molecular structure at the binding site.Isoenzymes: Structurally related forms of an enzyme. Each isoenzyme has the same mechanism and classification, but differs in its chemical, physical, or immunological characteristics.Molecular Weight: The sum of the weight of all the atoms in a molecule.Electrophoresis, Polyacrylamide Gel: Electrophoresis in which a polyacrylamide gel is used as the diffusion medium.Molecular Structure: The location of the atoms, groups or ions relative to one another in a molecule, as well as the number, type and location of covalent bonds.Amino Acids: Organic compounds that generally contain an amino (-NH2) and a carboxyl (-COOH) group. Twenty alpha-amino acids are the subunits which are polymerized to form proteins.Peptides: Members of the class of compounds composed of AMINO ACIDS joined together by peptide bonds between adjacent amino acids into linear, branched or cyclical structures. OLIGOPEPTIDES are composed of approximately 2-12 amino acids. Polypeptides are composed of approximately 13 or more amino acids. PROTEINS are linear polypeptides that are normally synthesized on RIBOSOMES.Enzyme Stability: The extent to which an enzyme retains its structural conformation or its activity when subjected to storage, isolation, and purification or various other physical or chemical manipulations, including proteolytic enzymes and heat.Phosphorylation: The introduction of a phosphoryl group into a compound through the formation of an ester bond between the compound and a phosphorus moiety.Amino Acid Substitution: The naturally occurring or experimentally induced replacement of one or more AMINO ACIDS in a protein with another. If a functionally equivalent amino acid is substituted, the protein may retain wild-type activity. Substitution may also diminish, enhance, or eliminate protein function. Experimentally induced substitution is often used to study enzyme activities and binding site properties.Oligopeptides: Peptides composed of between two and twelve amino acids.Stereoisomerism: The phenomenon whereby compounds whose molecules have the same number and kind of atoms and the same atomic arrangement, but differ in their spatial relationships. (From McGraw-Hill Dictionary of Scientific and Technical Terms, 5th ed)Temperature: The property of objects that determines the direction of heat flow when they are placed in direct thermal contact. The temperature is the energy of microscopic motions (vibrational and translational) of the particles of atoms.Recombinant Fusion Proteins: Recombinant proteins produced by the GENETIC TRANSLATION of fused genes formed by the combination of NUCLEIC ACID REGULATORY SEQUENCES of one or more genes with the protein coding sequences of one or more genes.Cell Line: Established cell cultures that have the potential to propagate indefinitely.DNA Primers: Short sequences (generally about 10 base pairs) of DNA that are complementary to sequences of messenger RNA and allow reverse transcriptases to start copying the adjacent sequences of mRNA. Primers are used extensively in genetic and molecular biology techniques.Endopeptidases: A subclass of PEPTIDE HYDROLASES that catalyze the internal cleavage of PEPTIDES or PROTEINS.Serine Endopeptidases: Any member of the group of ENDOPEPTIDASES containing at the active site a serine residue involved in catalysis.Chromatography, High Pressure Liquid: Liquid chromatographic techniques which feature high inlet pressures, high sensitivity, and high speed.Alcohol Oxidoreductases: A subclass of enzymes which includes all dehydrogenases acting on primary and secondary alcohols as well as hemiacetals. They are further classified according to the acceptor which can be NAD+ or NADP+ (subclass 1.1.1), cytochrome (1.1.2), oxygen (1.1.3), quinone (1.1.5), or another acceptor (1.1.99).Protein Structure, Secondary: The level of protein structure in which regular hydrogen-bond interactions within contiguous stretches of polypeptide chain give rise to alpha helices, beta strands (which align to form beta sheets) or other types of coils. This is the first folding level of protein conformation.Carbohydrate Sequence: The sequence of carbohydrates within POLYSACCHARIDES; GLYCOPROTEINS; and GLYCOLIPIDS.Saccharomyces cerevisiae: A species of the genus SACCHAROMYCES, family Saccharomycetaceae, order Saccharomycetales, known as "baker's" or "brewer's" yeast. The dried form is used as a dietary supplement.Escherichia coli Proteins: Proteins obtained from ESCHERICHIA COLI.Cattle: Domesticated bovine animals of the genus Bos, usually kept on a farm or ranch and used for the production of meat or dairy products or for heavy labor.Protein Engineering: Procedures by which protein structure and function are changed or created in vitro by altering existing or synthesizing new structural genes that direct the synthesis of proteins with sought-after properties. Such procedures may include the design of MOLECULAR MODELS of proteins using COMPUTER GRAPHICS or other molecular modeling techniques; site-specific mutagenesis (MUTAGENESIS, SITE-SPECIFIC) of existing genes; and DIRECTED MOLECULAR EVOLUTION techniques to create new genes.Phylogeny: The relationships of groups of organisms as reflected by their genetic makeup.Enzyme Activation: Conversion of an inactive form of an enzyme to one possessing metabolic activity. It includes 1, activation by ions (activators); 2, activation by cofactors (coenzymes); and 3, conversion of an enzyme precursor (proenzyme or zymogen) to an active enzyme.Oxidation-Reduction: A chemical reaction in which an electron is transferred from one molecule to another. The electron-donating molecule is the reducing agent or reductant; the electron-accepting molecule is the oxidizing agent or oxidant. Reducing and oxidizing agents function as conjugate reductant-oxidant pairs or redox pairs (Lehninger, Principles of Biochemistry, 1982, p471).Models, Chemical: Theoretical representations that simulate the behavior or activity of chemical processes or phenomena; includes the use of mathematical equations, computers, and other electronic equipment.Acyltransferases: Enzymes from the transferase class that catalyze the transfer of acyl groups from donor to acceptor, forming either esters or amides. (From Enzyme Nomenclature 1992) EC 2.3.Chromatography, Gel: Chromatography on non-ionic gels without regard to the mechanism of solute discrimination.Amino Acid Motifs: Commonly observed structural components of proteins formed by simple combinations of adjacent secondary structures. A commonly observed structure may be composed of a CONSERVED SEQUENCE which can be represented by a CONSENSUS SEQUENCE.DNA, Complementary: Single-stranded complementary DNA synthesized from an RNA template by the action of RNA-dependent DNA polymerase. cDNA (i.e., complementary DNA, not circular DNA, not C-DNA) is used in a variety of molecular cloning experiments as well as serving as a specific hybridization probe.Peptide Library: A collection of cloned peptides, or chemically synthesized peptides, frequently consisting of all possible combinations of amino acids making up an n-amino acid peptide.Glycoside HydrolasesDNA: A deoxyribonucleotide polymer that is the primary genetic material of all cells. Eukaryotic and prokaryotic organisms normally contain DNA in a double-stranded state, yet several important biological processes transiently involve single-stranded regions. DNA, which consists of a polysugar-phosphate backbone possessing projections of purines (adenine and guanine) and pyrimidines (thymine and cytosine), forms a double helix that is held together by hydrogen bonds between these purines and pyrimidines (adenine to thymine and guanine to cytosine).Oligosaccharides: Carbohydrates consisting of between two (DISACCHARIDES) and ten MONOSACCHARIDES connected by either an alpha- or beta-glycosidic link. They are found throughout nature in both the free and bound form.Liver: A large lobed glandular organ in the abdomen of vertebrates that is responsible for detoxification, metabolism, synthesis and storage of various substances.Biological Transport: The movement of materials (including biochemical substances and drugs) through a biological system at the cellular level. The transport can be across cell membranes and epithelial layers. It also can occur within intracellular compartments and extracellular compartments.Oxidoreductases: The class of all enzymes catalyzing oxidoreduction reactions. The substrate that is oxidized is regarded as a hydrogen donor. The systematic name is based on donor:acceptor oxidoreductase. The recommended name will be dehydrogenase, wherever this is possible; as an alternative, reductase can be used. Oxidase is only used in cases where O2 is the acceptor. (Enzyme Nomenclature, 1992, p9)Hydrolases: Any member of the class of enzymes that catalyze the cleavage of the substrate and the addition of water to the resulting molecules, e.g., ESTERASES, glycosidases (GLYCOSIDE HYDROLASES), lipases, NUCLEOTIDASES, peptidases (PEPTIDE HYDROLASES), and phosphatases (PHOSPHORIC MONOESTER HYDROLASES). EC 3.Carrier Proteins: Transport proteins that carry specific substances in the blood or across cell membranes.Adenosine Triphosphate: An adenine nucleotide containing three phosphate groups esterified to the sugar moiety. In addition to its crucial roles in metabolism adenosine triphosphate is a neurotransmitter.Plasmids: Extrachromosomal, usually CIRCULAR DNA molecules that are self-replicating and transferable from one organism to another. They are found in a variety of bacterial, archaeal, fungal, algal, and plant species. They are used in GENETIC ENGINEERING as CLONING VECTORS.Acyl Coenzyme A: S-Acyl coenzyme A. Fatty acid coenzyme A derivatives that are involved in the biosynthesis and oxidation of fatty acids as well as in ceramide formation.Biocatalysis: The facilitation of biochemical reactions with the aid of naturally occurring catalysts such as ENZYMES.Membrane Transport Proteins: Membrane proteins whose primary function is to facilitate the transport of molecules across a biological membrane. Included in this broad category are proteins involved in active transport (BIOLOGICAL TRANSPORT, ACTIVE), facilitated transport and ION CHANNELS.Peptide Hydrolases: Hydrolases that specifically cleave the peptide bonds found in PROTEINS and PEPTIDES. Examples of sub-subclasses for this group include EXOPEPTIDASES and ENDOPEPTIDASES.Protein Kinases: A family of enzymes that catalyze the conversion of ATP and a protein to ADP and a phosphoprotein.Conserved Sequence: A sequence of amino acids in a polypeptide or of nucleotides in DNA or RNA that is similar across multiple species. A known set of conserved sequences is represented by a CONSENSUS SEQUENCE. AMINO ACID MOTIFS are often composed of conserved sequences.Aminopeptidases: A subclass of EXOPEPTIDASES that act on the free N terminus end of a polypeptide liberating a single amino acid residue. EC 3.4.11.beta-Glucosidase: An exocellulase with specificity for a variety of beta-D-glycoside substrates. It catalyzes the hydrolysis of terminal non-reducing residues in beta-D-glucosides with release of GLUCOSE.Pseudomonas: A genus of gram-negative, aerobic, rod-shaped bacteria widely distributed in nature. Some species are pathogenic for humans, animals, and plants.Mixed Function Oxygenases: Widely distributed enzymes that carry out oxidation-reduction reactions in which one atom of the oxygen molecule is incorporated into the organic substrate; the other oxygen atom is reduced and combined with hydrogen ions to form water. They are also known as monooxygenases or hydroxylases. These reactions require two substrates as reductants for each of the two oxygen atoms. There are different classes of monooxygenases depending on the type of hydrogen-providing cosubstrate (COENZYMES) required in the mixed-function oxidation.Carboxylic Ester Hydrolases: Enzymes which catalyze the hydrolysis of carboxylic acid esters with the formation of an alcohol and a carboxylic acid anion.Mutagenesis: Process of generating a genetic MUTATION. It may occur spontaneously or be induced by MUTAGENS.Peptide Fragments: Partial proteins formed by partial hydrolysis of complete proteins or generated through PROTEIN ENGINEERING techniques.Enzyme Inhibitors: Compounds or agents that combine with an enzyme in such a manner as to prevent the normal substrate-enzyme combination and the catalytic reaction.Dioxygenases: Non-heme iron-containing enzymes that incorporate two atoms of OXYGEN into the substrate. They are important in biosynthesis of FLAVONOIDS; GIBBERELLINS; and HYOSCYAMINE; and for degradation of AROMATIC HYDROCARBONS.Cations, Divalent: Positively charged atoms, radicals or groups of atoms with a valence of plus 2, which travel to the cathode or negative pole during electrolysis.Cysteine Endopeptidases: ENDOPEPTIDASES which have a cysteine involved in the catalytic process. This group of enzymes is inactivated by CYSTEINE PROTEINASE INHIBITORS such as CYSTATINS and SULFHYDRYL REAGENTS.Membrane Proteins: Proteins which are found in membranes including cellular and intracellular membranes. They consist of two types, peripheral and integral proteins. They include most membrane-associated enzymes, antigenic proteins, transport proteins, and drug, hormone, and lectin receptors.Protease Inhibitors: Compounds which inhibit or antagonize biosynthesis or actions of proteases (ENDOPEPTIDASES).AmidohydrolasesRabbits: The species Oryctolagus cuniculus, in the family Leporidae, order LAGOMORPHA. Rabbits are born in burrows, furless, and with eyes and ears closed. In contrast with HARES, rabbits have 22 chromosome pairs.Binding, Competitive: The interaction of two or more substrates or ligands with the same binding site. The displacement of one by the other is used in quantitative and selective affinity measurements.Models, Biological: Theoretical representations that simulate the behavior or activity of biological processes or diseases. For disease models in living animals, DISEASE MODELS, ANIMAL is available. Biological models include the use of mathematical equations, computers, and other electronic equipment.Saccharomyces cerevisiae Proteins: Proteins obtained from the species SACCHAROMYCES CEREVISIAE. The function of specific proteins from this organism are the subject of intense scientific interest and have been used to derive basic understanding of the functioning similar proteins in higher eukaryotes.Alanine: A non-essential amino acid that occurs in high levels in its free state in plasma. It is produced from pyruvate by transamination. It is involved in sugar and acid metabolism, increases IMMUNITY, and provides energy for muscle tissue, BRAIN, and the CENTRAL NERVOUS SYSTEM.Mass Spectrometry: An analytical method used in determining the identity of a chemical based on its mass using mass analyzers/mass spectrometers.EsterasesStreptomyces: A genus of bacteria that form a nonfragmented aerial mycelium. Many species have been identified with some being pathogenic. This genus is responsible for producing a majority of the ANTI-BACTERIAL AGENTS of practical value.Peptide Synthases: Ligases that catalyze the joining of adjacent AMINO ACIDS by the formation of carbon-nitrogen bonds between their carboxylic acid groups and amine groups.Organ Specificity: Characteristic restricted to a particular organ of the body, such as a cell type, metabolic response or expression of a particular protein or antigen.Chromatography: Techniques used to separate mixtures of substances based on differences in the relative affinities of the substances for mobile and stationary phases. A mobile phase (fluid or gas) passes through a column containing a stationary phase of porous solid or liquid coated on a solid support. Usage is both analytical for small amounts and preparative for bulk amounts.Dipeptides: Peptides composed of two amino acid units.Phosphoric Monoester Hydrolases: A group of hydrolases which catalyze the hydrolysis of monophosphoric esters with the production of one mole of orthophosphate. EC 3.1.3.Cell Membrane: The lipid- and protein-containing, selectively permeable membrane that surrounds the cytoplasm in prokaryotic and eukaryotic cells.Metals: Electropositive chemical elements characterized by ductility, malleability, luster, and conductance of heat and electricity. They can replace the hydrogen of an acid and form bases with hydroxyl radicals. (Grant & Hackh's Chemical Dictionary, 5th ed)Arginine: An essential amino acid that is physiologically active in the L-form.Time Factors: Elements of limited time intervals, contributing to particular results or situations.Chromatography, Ion Exchange: Separation technique in which the stationary phase consists of ion exchange resins. The resins contain loosely held small ions that easily exchange places with other small ions of like charge present in solutions washed over the resins.Dimerization: The process by which two molecules of the same chemical composition form a condensation product or polymer.NAD: A coenzyme composed of ribosylnicotinamide 5'-diphosphate coupled to adenosine 5'-phosphate by pyrophosphate linkage. It is found widely in nature and is involved in numerous enzymatic reactions in which it serves as an electron carrier by being alternately oxidized (NAD+) and reduced (NADH). (Dorland, 27th ed)Magnetic Resonance Spectroscopy: Spectroscopic method of measuring the magnetic moment of elementary particles such as atomic nuclei, protons or electrons. It is employed in clinical applications such as NMR Tomography (MAGNETIC RESONANCE IMAGING).Sulfotransferases: Enzymes which transfer sulfate groups to various acceptor molecules. They are involved in posttranslational sulfation of proteins and sulfate conjugation of exogenous chemicals and bile acids. EC 2.8.2.Transfection: The uptake of naked or purified DNA by CELLS, usually meaning the process as it occurs in eukaryotic cells. It is analogous to bacterial transformation (TRANSFORMATION, BACTERIAL) and both are routinely employed in GENE TRANSFER TECHNIQUES.Cytochrome P-450 Enzyme System: A superfamily of hundreds of closely related HEMEPROTEINS found throughout the phylogenetic spectrum, from animals, plants, fungi, to bacteria. They include numerous complex monooxygenases (MIXED FUNCTION OXYGENASES). In animals, these P-450 enzymes serve two major functions: (1) biosynthesis of steroids, fatty acids, and bile acids; (2) metabolism of endogenous and a wide variety of exogenous substrates, such as toxins and drugs (BIOTRANSFORMATION). They are classified, according to their sequence similarities rather than functions, into CYP gene families (>40% homology) and subfamilies (>59% homology). For example, enzymes from the CYP1, CYP2, and CYP3 gene families are responsible for most drug metabolism.Lysine: An essential amino acid. It is often added to animal feed.Dipeptidases: EXOPEPTIDASES that specifically act on dipeptides. EC 3.4.13.Chromatography, Affinity: A chromatographic technique that utilizes the ability of biological molecules to bind to certain ligands specifically and reversibly. It is used in protein biochemistry. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)Polymerase Chain Reaction: In vitro method for producing large amounts of specific DNA or RNA fragments of defined length and sequence from small amounts of short oligonucleotide flanking sequences (primers). The essential steps include thermal denaturation of the double-stranded target molecules, annealing of the primers to their complementary sequences, and extension of the annealed primers by enzymatic synthesis with DNA polymerase. The reaction is efficient, specific, and extremely sensitive. Uses for the reaction include disease diagnosis, detection of difficult-to-isolate pathogens, mutation analysis, genetic testing, DNA sequencing, and analyzing evolutionary relationships.Bacillus: A genus of BACILLACEAE that are spore-forming, rod-shaped cells. Most species are saprophytic soil forms with only a few species being pathogenic.Metalloendopeptidases: ENDOPEPTIDASES which use a metal such as ZINC in the catalytic mechanism.Pichia: Yeast-like ascomycetous fungi of the family Saccharomycetaceae, order SACCHAROMYCETALES isolated from exuded tree sap.Structural Homology, Protein: The degree of 3-dimensional shape similarity between proteins. It can be an indication of distant AMINO ACID SEQUENCE HOMOLOGY and used for rational DRUG DESIGN.Aspartic Acid: One of the non-essential amino acids commonly occurring in the L-form. It is found in animals and plants, especially in sugar cane and sugar beets. It may be a neurotransmitter.Protein Processing, Post-Translational: Any of various enzymatically catalyzed post-translational modifications of PEPTIDES or PROTEINS in the cell of origin. These modifications include carboxylation; HYDROXYLATION; ACETYLATION; PHOSPHORYLATION; METHYLATION; GLYCOSYLATION; ubiquitination; oxidation; proteolysis; and crosslinking and result in changes in molecular weight and electrophoretic motility.Serine: A non-essential amino acid occurring in natural form as the L-isomer. It is synthesized from GLYCINE or THREONINE. It is involved in the biosynthesis of PURINES; PYRIMIDINES; and other amino acids.PolysaccharidesAdenosine Triphosphatases: A group of enzymes which catalyze the hydrolysis of ATP. The hydrolysis reaction is usually coupled with another function such as transporting Ca(2+) across a membrane. These enzymes may be dependent on Ca(2+), Mg(2+), anions, H+, or DNA.Disaccharides: Oligosaccharides containing two monosaccharide units linked by a glycosidic bond.Coumarins: Synthetic or naturally occurring substances related to coumarin, the delta-lactone of coumarinic acid.Chromatography, Thin Layer: Chromatography on thin layers of adsorbents rather than in columns. The adsorbent can be alumina, silica gel, silicates, charcoals, or cellulose. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)Oxygenases: Oxidases that specifically introduce DIOXYGEN-derived oxygen atoms into a variety of organic molecules.HeLa Cells: The first continuously cultured human malignant CELL LINE, derived from the cervical carcinoma of Henrietta Lacks. These cells are used for VIRUS CULTIVATION and antitumor drug screening assays.Molecular Conformation: The characteristic three-dimensional shape of a molecule.Cells, Cultured: Cells propagated in vitro in special media conducive to their growth. Cultured cells are used to study developmental, morphologic, metabolic, physiologic, and genetic processes, among others.Glutathione Transferase: A transferase that catalyzes the addition of aliphatic, aromatic, or heterocyclic FREE RADICALS as well as EPOXIDES and arene oxides to GLUTATHIONE. Addition takes place at the SULFUR. It also catalyzes the reduction of polyol nitrate by glutathione to polyol and nitrite.Multigene Family: A set of genes descended by duplication and variation from some ancestral gene. Such genes may be clustered together on the same chromosome or dispersed on different chromosomes. Examples of multigene families include those that encode the hemoglobins, immunoglobulins, histocompatibility antigens, actins, tubulins, keratins, collagens, heat shock proteins, salivary glue proteins, chorion proteins, cuticle proteins, yolk proteins, and phaseolins, as well as histones, ribosomal RNA, and transfer RNA genes. The latter three are examples of reiterated genes, where hundreds of identical genes are present in a tandem array. (King & Stanfield, A Dictionary of Genetics, 4th ed)Pseudomonas putida: A species of gram-negative, aerobic bacteria isolated from soil and water as well as clinical specimens. Occasionally it is an opportunistic pathogen.Sequence Analysis, DNA: A multistage process that includes cloning, physical mapping, subcloning, determination of the DNA SEQUENCE, and information analysis.Enzyme Assays: Methods used to measure the relative activity of a specific enzyme or its concentration in solution. Typically an enzyme substrate is added to a buffer solution containing enzyme and the rate of conversion of substrate to product is measured under controlled conditions. Many classical enzymatic assay methods involve the use of synthetic colorimetric substrates and measuring the reaction rates using a spectrophotometer.N-Acetylgalactosaminyltransferases: Enzymes that catalyze the transfer of N-acetylgalactosamine from a nucleoside diphosphate N-acetylgalactosamine to an acceptor molecule which is frequently another carbohydrate. EC 2.4.1.-.Manganese: A trace element with atomic symbol Mn, atomic number 25, and atomic weight 54.94. It is concentrated in cell mitochondria, mostly in the pituitary gland, liver, pancreas, kidney, and bone, influences the synthesis of mucopolysaccharides, stimulates hepatic synthesis of cholesterol and fatty acids, and is a cofactor in many enzymes, including arginase and alkaline phosphatase in the liver. (From AMA Drug Evaluations Annual 1992, p2035)Methyltransferases: A subclass of enzymes of the transferase class that catalyze the transfer of a methyl group from one compound to another. (Dorland, 28th ed) EC 2.1.1.Protein Folding: Processes involved in the formation of TERTIARY PROTEIN STRUCTURE.Carboxypeptidases: Enzymes that act at a free C-terminus of a polypeptide to liberate a single amino acid residue.Protein Structure, Quaternary: The characteristic 3-dimensional shape and arrangement of multimeric proteins (aggregates of more than one polypeptide chain).Hydroxylation: Placing of a hydroxyl group on a compound in a position where one did not exist before. (Stedman, 26th ed)Aspergillus niger: An imperfect fungus causing smut or black mold of several fruits, vegetables, etc.Swine: Any of various animals that constitute the family Suidae and comprise stout-bodied, short-legged omnivorous mammals with thick skin, usually covered with coarse bristles, a rather long mobile snout, and small tail. Included are the genera Babyrousa, Phacochoerus (wart hogs), and Sus, the latter containing the domestic pig (see SUS SCROFA).Fungal Proteins: Proteins found in any species of fungus.Ligands: A molecule that binds to another molecule, used especially to refer to a small molecule that binds specifically to a larger molecule, e.g., an antigen binding to an antibody, a hormone or neurotransmitter binding to a receptor, or a substrate or allosteric effector binding to an enzyme. Ligands are also molecules that donate or accept a pair of electrons to form a coordinate covalent bond with the central metal atom of a coordination complex. (From Dorland, 27th ed)Carbohydrate Conformation: The characteristic 3-dimensional shape of a carbohydrate.Proteins: Linear POLYPEPTIDES that are synthesized on RIBOSOMES and may be further modified, crosslinked, cleaved, or assembled into complex proteins with several subunits. The specific sequence of AMINO ACIDS determines the shape the polypeptide will take, during PROTEIN FOLDING, and the function of the protein.Glycosylation: The chemical or biochemical addition of carbohydrate or glycosyl groups to other chemicals, especially peptides or proteins. Glycosyl transferases are used in this biochemical reaction.Tyrosine: A non-essential amino acid. In animals it is synthesized from PHENYLALANINE. It is also the precursor of EPINEPHRINE; THYROID HORMONES; and melanin.Mannosidases: Glycoside hydrolases that catalyze the hydrolysis of alpha or beta linked MANNOSE.COS Cells: CELL LINES derived from the CV-1 cell line by transformation with a replication origin defective mutant of SV40 VIRUS, which codes for wild type large T antigen (ANTIGENS, POLYOMAVIRUS TRANSFORMING). They are used for transfection and cloning. (The CV-1 cell line was derived from the kidney of an adult male African green monkey (CERCOPITHECUS AETHIOPS).)Chromatography, DEAE-Cellulose: A type of ion exchange chromatography using diethylaminoethyl cellulose (DEAE-CELLULOSE) as a positively charged resin. (From McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)Glucosidases: Enzymes that hydrolyze O-glucosyl-compounds. (Enzyme Nomenclature, 1992) EC 3.2.1.-.EstersCrystallization: The formation of crystalline substances from solutions or melts. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)Directed Molecular Evolution: The techniques used to produce molecules exhibiting properties that conform to the demands of the experimenter. These techniques combine methods of generating structural changes with methods of selection. They are also used to examine proposed mechanisms of evolution under in vitro selection conditions.Acetyltransferases: Enzymes catalyzing the transfer of an acetyl group, usually from acetyl coenzyme A, to another compound. EC 2.3.1.NADP: Nicotinamide adenine dinucleotide phosphate. A coenzyme composed of ribosylnicotinamide 5'-phosphate (NMN) coupled by pyrophosphate linkage to the 5'-phosphate adenosine 2',5'-bisphosphate. It serves as an electron carrier in a number of reactions, being alternately oxidized (NADP+) and reduced (NADPH). (Dorland, 27th ed)Cytosol: Intracellular fluid from the cytoplasm after removal of ORGANELLES and other insoluble cytoplasmic components.Nucleic Acid Conformation: The spatial arrangement of the atoms of a nucleic acid or polynucleotide that results in its characteristic 3-dimensional shape.Plant Proteins: Proteins found in plants (flowers, herbs, shrubs, trees, etc.). The concept does not include proteins found in vegetables for which VEGETABLE PROTEINS is available.Coenzyme A Ligases: Enzymes that catalyze the formation of acyl-CoA derivatives. EC 6.2.1.Transaminases: A subclass of enzymes of the transferase class that catalyze the transfer of an amino group from a donor (generally an amino acid) to an acceptor (generally a 2-keto acid). Most of these enzymes are pyridoxyl phosphate proteins. (Dorland, 28th ed) EC 2.6.1.Isoelectric Point: The pH in solutions of proteins and related compounds at which the dipolar ions are at a maximum.Glycine: A non-essential amino acid. It is found primarily in gelatin and silk fibroin and used therapeutically as a nutrient. It is also a fast inhibitory neurotransmitter.Thermodynamics: A rigorously mathematical analysis of energy relationships (heat, work, temperature, and equilibrium). It describes systems whose states are determined by thermal parameters, such as temperature, in addition to mechanical and electromagnetic parameters. (From Hawley's Condensed Chemical Dictionary, 12th ed)alpha-Mannosidase: An enzyme that catalyzes the HYDROLYSIS of terminal, non-reducing alpha-D-mannose residues in alpha-D-mannosides. The enzyme plays a role in the processing of newly formed N-glycans and in degradation of mature GLYCOPROTEINS. There are multiple isoforms of alpha-mannosidase, each having its own specific cellular location and pH optimum. Defects in the lysosomal form of the enzyme results in a buildup of mannoside intermediate metabolites and the disease ALPHA-MANNOSIDOSIS.Hydrogen Bonding: A low-energy attractive force between hydrogen and another element. It plays a major role in determining the properties of water, proteins, and other compounds.Hot Temperature: Presence of warmth or heat or a temperature notably higher than an accustomed norm.DNA, Bacterial: Deoxyribonucleic acid that makes up the genetic material of bacteria.Trypsin: A serine endopeptidase that is formed from TRYPSINOGEN in the pancreas. It is converted into its active form by ENTEROPEPTIDASE in the small intestine. It catalyzes hydrolysis of the carboxyl group of either arginine or lysine. EC 3.4.21.4.Alcohols: Alkyl compounds containing a hydroxyl group. They are classified according to relation of the carbon atom: primary alcohols, R-CH2OH; secondary alcohols, R2-CHOH; tertiary alcohols, R3-COH. (From Grant & Hackh's Chemical Dictionary, 5th ed)N-Glycosyl Hydrolases: A class of enzymes involved in the hydrolysis of the N-glycosidic bond of nitrogen-linked sugars.Gene Expression: The phenotypic manifestation of a gene or genes by the processes of GENETIC TRANSCRIPTION and GENETIC TRANSLATION.Genes, Bacterial: The functional hereditary units of BACTERIA.Evolution, Molecular: The process of cumulative change at the level of DNA; RNA; and PROTEINS, over successive generations.Lipase: An enzyme of the hydrolase class that catalyzes the reaction of triacylglycerol and water to yield diacylglycerol and a fatty acid anion. It is produced by glands on the tongue and by the pancreas and initiates the digestion of dietary fats. (From Dorland, 27th ed) EC 3.1.1.3.Microsomes, Liver: Closed vesicles of fragmented endoplasmic reticulum created when liver cells or tissue are disrupted by homogenization. They may be smooth or rough.Phenylalanine: An essential aromatic amino acid that is a precursor of MELANIN; DOPAMINE; noradrenalin (NOREPINEPHRINE), and THYROXINE.Signal Transduction: The intracellular transfer of information (biological activation/inhibition) through a signal pathway. In each signal transduction system, an activation/inhibition signal from a biologically active molecule (hormone, neurotransmitter) is mediated via the coupling of a receptor/enzyme to a second messenger system or to an ion channel. Signal transduction plays an important role in activating cellular functions, cell differentiation, and cell proliferation. Examples of signal transduction systems are the GAMMA-AMINOBUTYRIC ACID-postsynaptic receptor-calcium ion channel system, the receptor-mediated T-cell activation pathway, and the receptor-mediated activation of phospholipases. Those coupled to membrane depolarization or intracellular release of calcium include the receptor-mediated activation of cytotoxic functions in granulocytes and the synaptic potentiation of protein kinase activation. Some signal transduction pathways may be part of larger signal transduction pathways; for example, protein kinase activation is part of the platelet activation signal pathway.Blotting, Western: Identification of proteins or peptides that have been electrophoretically separated by blot transferring from the electrophoresis gel to strips of nitrocellulose paper, followed by labeling with antibody probes.Enzymes: Biological molecules that possess catalytic activity. They may occur naturally or be synthetically created. Enzymes are usually proteins, however CATALYTIC RNA and CATALYTIC DNA molecules have also been identified.Spectrophotometry: The art or process of comparing photometrically the relative intensities of the light in different parts of the spectrum.Reproducibility of Results: The statistical reproducibility of measurements (often in a clinical context), including the testing of instrumentation or techniques to obtain reproducible results. The concept includes reproducibility of physiological measurements, which may be used to develop rules to assess probability or prognosis, or response to a stimulus; reproducibility of occurrence of a condition; and reproducibility of experimental results.Subtilisins: A family of SERINE ENDOPEPTIDASES isolated from Bacillus subtilis. EC 3.4.21.-Cysteine: A thiol-containing non-essential amino acid that is oxidized to form CYSTINE.Plants: Multicellular, eukaryotic life forms of kingdom Plantae (sensu lato), comprising the VIRIDIPLANTAE; RHODOPHYTA; and GLAUCOPHYTA; all of which acquired chloroplasts by direct endosymbiosis of CYANOBACTERIA. They are characterized by a mainly photosynthetic mode of nutrition; essentially unlimited growth at localized regions of cell divisions (MERISTEMS); cellulose within cells providing rigidity; the absence of organs of locomotion; absence of nervous and sensory systems; and an alternation of haploid and diploid generations.Oligodeoxyribonucleotides: A group of deoxyribonucleotides (up to 12) in which the phosphate residues of each deoxyribonucleotide act as bridges in forming diester linkages between the deoxyribose moieties.Penicillin Amidase: An enzyme catalyzing the hydrolysis of penicillin to penicin and a carboxylic acid anion. EC 3.5.1.11.NitrophenolsRNA, Messenger: RNA sequences that serve as templates for protein synthesis. Bacterial mRNAs are generally primary transcripts in that they do not require post-transcriptional processing. Eukaryotic mRNA is synthesized in the nucleus and must be exported to the cytoplasm for translation. Most eukaryotic mRNAs have a sequence of polyadenylic acid at the 3' end, referred to as the poly(A) tail. The function of this tail is not known for certain, but it may play a role in the export of mature mRNA from the nucleus as well as in helping stabilize some mRNA molecules by retarding their degradation in the cytoplasm.Threonine: An essential amino acid occurring naturally in the L-form, which is the active form. It is found in eggs, milk, gelatin, and other proteins.Trisaccharides: Oligosaccharides containing three monosaccharide units linked by glycosidic bonds.Macromolecular Substances: Compounds and molecular complexes that consist of very large numbers of atoms and are generally over 500 kDa in size. In biological systems macromolecular substances usually can be visualized using ELECTRON MICROSCOPY and are distinguished from ORGANELLES by the lack of a membrane structure.Methylation: Addition of methyl groups. In histo-chemistry methylation is used to esterify carboxyl groups and remove sulfate groups by treating tissue sections with hot methanol in the presence of hydrochloric acid. (From Stedman, 25th ed)

Prodigious substrate specificity of AAC(6')-APH(2"), an aminoglycoside antibiotic resistance determinant in enterococci and staphylococci. (1/34326)

BACKGROUND: High-level gentamicin resistance in enterococci and staphylococci is conferred by AAC(6')-APH(2"), an enzyme with 6'-N-acetyltransferase and 2"-O-phosphotransferase activities. The presence of this enzyme in pathogenic gram-positive bacteria prevents the successful use of gentamicin C and most other aminoglycosides as therapeutic agents. RESULTS: In an effort to understand the mechanism of aminoglycoside modification, we expressed AAC(6')-APH(2") in Bacillus subtilis. The purified enzyme is monomeric with a molecular mass of 57 kDa and displays both the expected aminoglycoside N-acetyltransferase and O-phosphotransferase activities. Structure-function analysis with various aminoglycosides substrates reveals an enzyme with broad specificity in both enzymatic activities, accounting for AAC(6')-APH(2")'s dramatic negative impact on clinical aminoglycoside therapy. Both lividomycin A and paromomycin, aminoglycosides lacking a 6'-amino group, were acetylated by AAC(6')-APH(2"). The infrared spectrum of the product of paromomycin acetylation yielded a signal consistent with O-acetylation. Mass spectral and nuclear magnetic resonance analysis of the products of neomycin phosphorylation indicated that phosphoryl transfer occurred primarily at the 3'-OH of the 6-aminohexose ring A, and that some diphosphorylated material was also present with phosphates at the 3'-OH and the 3"'-OH of ring D, both unprecedented observations for this enzyme. Furthermore, the phosphorylation site of lividomycin A was determined to be the 5"-OH of the pentose ring C. CONCLUSIONS: The bifunctional AAC(6')-APH(2") has the capacity to inactivate virtually all clinically important aminoglycosides through N- and O-acetylation and phosphorylation of hydroxyl groups. The extremely broad substrate specificity of this enzyme will impact on future development of aminoglycosides and presents a significant challenge for antibiotic design.  (+info)

Mechanism and specificity of the terminal thioesterase domain from the erythromycin polyketide synthase. (2/34326)

BACKGROUND: Polyketides are important compounds with antibiotic and anticancer activities. Several modular polyketide synthases (PKSs) contain a terminal thioesterase (TE) domain probably responsible for the release and concomitant cyclization of the fully processed polyketide chain. Because the TE domain influences qualitative aspects of product formation by engineered PKSs, its mechanism and specificity are of considerable interest. RESULTS: The TE domain of the 6-deoxyerythronolide B synthase was overexpressed in Escherichia coli. When tested against a set of N-acetyl cysteamine thioesters the TE domain did not act as a cyclase, but showed significant hydrolytic specificity towards substrates that mimic important features of its natural substrate. Also the overall rate of polyketide chain release was strongly enhanced by a covalent connection between the TE domain and the terminal PKS module (by as much as 100-fold compared with separate TE and PKS 'domains'). CONCLUSIONS: The inability of the TE domain alone to catalyze cyclization suggests that macrocycle formation results from the combined action of the TE domain and a PKS module. The chain-length and stereochemical preferences of the TE domain might be relevant in the design and engineered biosynthesis of certain novel polyketides. Our results also suggest that the TE domain might loop back to catalyze the release of polyketide chains from both terminal and pre-terminal modules, which may explain the ability of certain naturally occurring PKSs, such as the picromycin synthase, to generate both 12-membered and 14-membered macrolide antibiotics.  (+info)

A genetic model of substrate deprivation therapy for a glycosphingolipid storage disorder. (3/34326)

Inherited defects in the degradation of glycosphingolipids (GSLs) cause a group of severe diseases known as GSL storage disorders. There are currently no effective treatments for the majority of these disorders. We have explored a new treatment paradigm, substrate deprivation therapy, by constructing a genetic model in mice. Sandhoff's disease mice, which abnormally accumulate GSLs, were bred with mice that were blocked in their synthesis of GSLs. The mice with simultaneous defects in GSL synthesis and degradation no longer accumulated GSLs, had improved neurologic function, and had a much longer life span. However, these mice eventually developed a late-onset neurologic disease because of accumulation of another class of substrate, oligosaccharides. The results support the validity of the substrate deprivation therapy and also highlight some limitations.  (+info)

Activation of Src in human breast tumor cell lines: elevated levels of phosphotyrosine phosphatase activity that preferentially recognizes the Src carboxy terminal negative regulatory tyrosine 530. (4/34326)

Elevated levels of Src kinase activity have been reported in a number of human cancers, including colon and breast cancer. We have analysed four human breast tumor cell lines that exhibit high levels of Src kinase activity, and have determined that these cell lines also exhibit a high level of a phosphotyrosine phosphatase activity that recognizes the Src carboxy-terminal P-Tyr530 negative regulatory site. Total Src kinase activity in these cell lines is elevated as much as 30-fold over activity in normal control cells and specific activity is elevated as much as 5.6-fold. When the breast tumor cells were grown in the presence of the tyrosine phosphatase inhibitor vanadate, Src kinase activity was reduced in all four breast tumor cell lines, suggesting that Src was being activated by a phosphatase which could recognize the Tyr530 negative regulatory site. In fractionated cell extracts from the breast tumor cells, we found elevated levels of a membrane associated tyrosine phosphatase activity that preferentially dephosphorylated a Src family carboxy-terminal phosphopeptide containing the regulatory tyrosine 530 site. Src was hypophosphorylated in vivo at tyrosine 530 in at least two of the tumor cell lines, further suggesting that Src was being activated by a phosphatase in these cells. In preliminary immunoprecipitation and antibody depletion experiments, we were unable to correlate the major portion of this phosphatase activity with several known phosphatases.  (+info)

Crystal structure of MHC class II-associated p41 Ii fragment bound to cathepsin L reveals the structural basis for differentiation between cathepsins L and S. (5/34326)

The lysosomal cysteine proteases cathepsins S and L play crucial roles in the degradation of the invariant chain during maturation of MHC class II molecules and antigen processing. The p41 form of the invariant chain includes a fragment which specifically inhibits cathepsin L but not S. The crystal structure of the p41 fragment, a homologue of the thyroglobulin type-1 domains, has been determined at 2.0 A resolution in complex with cathepsin L. The structure of the p41 fragment demonstrates a novel fold, consisting of two subdomains, each stabilized by disulfide bridges. The first subdomain is an alpha-helix-beta-strand arrangement, whereas the second subdomain has a predominantly beta-strand arrangement. The wedge shape and three-loop arrangement of the p41 fragment bound to the active site cleft of cathepsin L are reminiscent of the inhibitory edge of cystatins, thus demonstrating the first example of convergent evolution observed in cysteine protease inhibitors. However, the different fold of the p41 fragment results in additional contacts with the top of the R-domain of the enzymes, which defines the specificity-determining S2 and S1' substrate-binding sites. This enables inhibitors based on the thyroglobulin type-1 domain fold, in contrast to the rather non-selective cystatins, to exhibit specificity for their target enzymes.  (+info)

Substrate specificities of SR proteins in constitutive splicing are determined by their RNA recognition motifs and composite pre-mRNA exonic elements. (6/34326)

We report striking differences in the substrate specificities of two human SR proteins, SF2/ASF and SC35, in constitutive splicing. beta-Globin pre-mRNA (exons 1 and 2) is spliced indiscriminately with either SR protein. Human immunodeficiency virus tat pre-mRNA (exons 2 and 3) and immunoglobulin mu-chain (IgM) pre-mRNA (exons C3 and C4) are preferentially spliced with SF2/ASF and SC35, respectively. Using in vitro splicing with mutated or chimeric derivatives of the tat and IgM pre-mRNAs, we defined specific combinations of segments in the downstream exons, which mediate either positive or negative effects to confer SR protein specificity. A series of recombinant chimeric proteins consisting of domains of SF2/ASF and SC35 in various combinations was used to localize trans-acting domains responsible for substrate specificity. The RS domains of SF2/ASF and SC35 can be exchanged without effect on substrate specificity. The RNA recognition motifs (RRMs) of SF2/ASF are active only in the context of a two-RRM structure, and RRM2 has a dominant role in substrate specificity. In contrast, the single RRM of SC35 can function alone, but its substrate specificity can be influenced by the presence of an additional RRM. The RRMs behave as modules that, when present in different combinations, can have positive, neutral, or negative effects on splicing, depending upon the specific substrate. We conclude that SR protein-specific recognition of specific positive and negative pre-mRNA exonic elements via one or more RRMs is a crucial determinant of the substrate specificity of SR proteins in constitutive splicing.  (+info)

Pseudouridine mapping in the Saccharomyces cerevisiae spliceosomal U small nuclear RNAs (snRNAs) reveals that pseudouridine synthase pus1p exhibits a dual substrate specificity for U2 snRNA and tRNA. (7/34326)

Pseudouridine (Psi) residues were localized in the Saccharomyces cerevisiae spliceosomal U small nuclear RNAs (UsnRNAs) by using the chemical mapping method. In contrast to vertebrate UsnRNAs, S. cerevisiae UsnRNAs contain only a few Psi residues, which are located in segments involved in intermolecular RNA-RNA or RNA-protein interactions. At these positions, UsnRNAs are universally modified. When yeast mutants disrupted for one of the several pseudouridine synthase genes (PUS1, PUS2, PUS3, and PUS4) or depleted in rRNA-pseudouridine synthase Cbf5p were tested for UsnRNA Psi content, only the loss of the Pus1p activity was found to affect Psi formation in spliceosomal UsnRNAs. Indeed, Psi44 formation in U2 snRNA was abolished. By using purified Pus1p enzyme and in vitro-produced U2 snRNA, Pus1p is shown here to catalyze Psi44 formation in the S. cerevisiae U2 snRNA. Thus, Pus1p is the first UsnRNA pseudouridine synthase characterized so far which exhibits a dual substrate specificity, acting on both tRNAs and U2 snRNA. As depletion of rRNA-pseudouridine synthase Cbf5p had no effect on UsnRNA Psi content, formation of Psi residues in S. cerevisiae UsnRNAs is not dependent on the Cbf5p-snoRNA guided mechanism.  (+info)

The human F box protein beta-Trcp associates with the Cul1/Skp1 complex and regulates the stability of beta-catenin. (8/34326)

Ubiquitin-conjugation targets numerous cellular regulators for proteasome-mediated degradation. Thus, the identification of ubiquitin ligases and their physiological substrates is crucially important, especially for those cases in which aberrant levels of regulatory proteins (e.g., beta-catenin, p27) result from a deregulated ubiquitination pathway. In yeast, the proteolysis of several G1 regulators is controlled by ubiquitin ligases (or SCFs) formed by three subunits: Skp1, Cul A (Cdc53), and one of many F-box proteins. Specific F-box proteins (Fbps) recruit different substrates to the SCF. Although many Fbps have been identified in mammals, their specific substrates and the existence of multiple SCFs have not yet been reported. We have found that one human Fbp, beta-Trcp (beta-Transducin repeat containing protein), does indeed form a novel SCF with human Skp1 and Cul1. Consistent with recent reports indicating that Xenopus and Drosophila beta-Trcp homologs act as negative regulators of the Wnt/beta-catenin signaling pathway, we report here that human beta-Trcp interacts with beta-catenin in vivo. Furthermore, beta-catenin is specifically stabilized in vivo by the expression of a dominant negative beta-Trcp. These results indicate that the Cul1/Skp1/beta-Trcp complex forms a ubiquitin ligase that mediates the degradation of beta-catenin.  (+info)

The present invention relates to methods for the conversion of the substrate specificity of desaturases. Specifically, the present invention pertains to a method for the conversion of the substrate specificity of a Δ5 and/or Δ6 desaturase to the substrate specificity of a Δ4 desaturase, the method comprising: identifying regions and/or amino acid residues which control the substrate specificity of (i) the Δ5 and/or Δ6 desaturase and (ii) the Δ4 desaturase; and replacing in the amino acid sequence of the mentioned Δ5 and/or Δ6 desaturase, the regions and/or amino acid residues which control the substrate specificity of the Δ5 and/or Δ6 desaturase, by the corresponding regions and/or amino acid residues which control the substrate specificity of the Δ4 desaturase, thereby converting the substrate specificity of the Δ5 and/or Δ6 desaturase to the substrate specificity of the Δ4 desaturase. The present invention further concerns a method for the conversion of the substrate specificity of a Δ4
Use:. This fluorescent ADAM substrate was originally described by us in the publication, " Fluorescent substrates useful as high-throughput screening tools for ADAM9″. This ADAM substrate is based on the cleavage sequence of precursor TNF-alpha and has been used to assess activity of ADAM17 in single cell assays as well as standard in vitro enzymatic and cell based assays (See publications below). See our Product Sheets for the substrate specificity profile of this substrate. This ADAM substrate is also an excellent substrate for ADAM9 and ADAM10. This substrate is not specific for ADAM family members as it can also be processed by members of the MMP family of proteinases. BioZyme Inc, does sell specific substrates for ADAM or MMP family members (Please see our Product Sheets or Catalog, for the substrate specificity profile). It demonstrates reasonably strong activity against all of those enzymes, with specificity constants, kcat/Km (M-1s-1), ranging from approximately 4 x 103 to 4 x 105. ...
Results presented in this report show that caspase activation after TCR triggering is a physiological, tightly regulated, and early response that appears to be required for efficient T cell activation. Indeed, the selective processing of caspase-3, -6, -7, and -8 was detected within 24 h after anti-CD3 stimulation of peripheral blood lymphocytes. Caspase processing occurred in various T and B cell subsets, and was found in proliferating and nonapoptotic lymphocytes. Activation of caspases was confirmed through binding of caspase-3-processed forms to a specific substrate, and by showing that a cell-permeable substrate was cleaved in intact, activated lymphocytes. Importantly, activation of the caspase cascade was associated to restricted substrate specificity, with cleavage of PARP and Wee1 being observed while two other substrates, DFF45 and RFC140, remained unaffected. Caspase processing after T cell stimulation correlated with a defective lymphocyte activation in the presence of the caspase ...
Steric and hydrophobic effects on substrate specificity were probed by protein engineering of subtilisin. Subtilisin has broad peptidase specificity and contains a large hydrophobic substrate binding cleft. A conserved glycine (Gly166), located at the bottom of the substrate binding left, was replaced by 12 nonionic amino acids by the cassette mutagenesis method. Mutant enzymes showed large changes in specificity toward substrates of increasing size and hydrophobicity. In general, the catalytic efficiency (kcat/Km) toward small hydrophobic substrates was increased (up to 16 times) by hydrophobic substitutions at position 166 in the binding cleft. Exceeding the optimal binding volume of the cleft (∼160 Å3), by enlarging either the substrate side chain or the side chain at position 166, evoked precipitous drops in catalytic efficiency (kcat/Km) (up to 5000 times) as a result of steric hindrance. ...
Protease-substrate interactions are governed by a variety of structural features. Although the substrate sequence specificities of numerous proteases have been established, topological specificities, whereby proteases may be classified based on recognition of distinct three-dimensional structural motifs, have not. The aggrecanase members of the ADAMTS (a disintegrin and metalloproteinase with thrombospondin motifs) family cleave a variety of proteins but do not seem to possess distinct sequence specificities. In the present study, the topological substrate specificity of ADAMTS-4 (aggrecanase-1) was examined using triple-helical or single-stranded poly(Pro) II helical peptides. Substrate topology modulated the affinity and sequence specificity of ADAMTS-4 with K(m) values indicating a preference for triple-helical structure. In turn, non-catalytic ADAMTS-4 domains were critical for hydrolysis of triple-helical and poly(Pro) II helical substrates. Comparison of ADAMTS-4 with MMP-1 (collagenase 1), MMP
X-converting enzyme (XCE) involved in nervous control of respiration, is a member of the M13 family of zinc peptidases, for which no natural substrate has been identified yet. In contrast, its well characterized homologue endothelin-converting enzyme-1 (ECE-1) showed broad substrate specificity and acts as endopeptidase as well as dipeptidase. To explore the structural differences between XCE and ECE-1, homology model of XCE was built using the complex structure of ECE-1 with phosphoramidon (pdb-id: 3DWB) as template. Phosphoramidon was docked into the binding site of XCE whereas phosphate oxygen of the inhibitor was used as water molecule to design the apo forms of both enzymes. Molecular dynamics simulation of both enzymes was performed to analyze the dynamic nature of their active site residues in the absence and presence of the inhibitor. Homology model of XCE explained the role of non-conserved residues of its S2 subsite. Molecular dynamics (MD) simulations identified the flexible transitions of
An electronic device may include first, second, and third substrates wherein the second electronic substrate is between the first and second electronic substrates. A first electrical and mechanical connection may be provided between the first and third electronic substrates, and a second electrical and mechanical connection may be provided between the second and third electronic substrates. In addition or in an alternative, an electronic device may include a printed circuit board, a first electronic substrate on the printed circuit board, a second electronic substrate on the first electronic substrate, and a third electronic substrate on the second electronic substrate. More particularly, the first electronic substrate may be between the printed circuit board and the second electronic substrate, and the second electronic substrate may be between the first and third electronic substrates. In addition, the second electronic substrate may be offset relative to the first and third electronic substrates so
UGT8 has been an "outlier" member of the UGT superfamily since its discovery in 1993 (Schulte and Stoffel, 1993). Initially called ceramide galactosyl-transferase, the gene was identified by purification of the protein conferring ceramide-galactosyl-transferase activity and protein sequencing, followed by screening of a rat brain cDNA library with probes derived from the deduced nucleotide sequence (Schulte and Stoffel, 1993). In common with other UGTs, UGT8 has a molecular mass in the 50-60 kDa range and carries the UGT signature sequence and motifs associated with retention in the endoplasmic reticulum membrane. Until now the substrate specificity of UGT8 has never been broadly investigated. We found that UGT8 had restricted substrate specificity and did not conjugate classic xenobiotic substrates common to most UGT1, 2, and 3 isoforms such as 4-methylumbelliferone and 4-nitrophenol (Uchaipichat et al., 2004), although it did conjugate one of the tested bioflavones (chrysin). The potent ...
We previously reported that the in vitro inhibitory effects of several OATP1B1 inhibitors showed remarkable substrate-dependence using prototypical substrates, E2G, E1S, and BSP (Izumi et al., 2013). In addition to the prototypical substrates, clinically used OATP1B1 substrate drugs could also serve as in vitro OATP1B1 probe substrates, for which the potential substrate-dependent inhibition has not been comprehensively evaluated. To identify representative in vitro OATP1B1 probe substrates that could mitigate the risk of false-negative DDI prediction, this study investigated the impact of in vitro substrate selection on OATP1B1 inhibition and the subsequent DDI prediction for 12 clinically used OATP1B1 substrate drugs compared with the prototypical probe substrates.. Twelve OATP1B1 substrate drugs-including statins (pitavastatin, atorvastatin, fluvastatin, rosuvastatin, and pravastatin), antidiabetics (repaglinide, nateglinide, and glibenclamide), a dual endothelin receptor antagonist ...
Cytochrome P450 (CYP) enzymes represent a large superfamily that displays extraordinarily diverse substrate specificities. After a concise review about CYPs of the CYP1A subfamily, which plays a crucial role in procarcinogen activation, this paper presents segment-directed mutagenesis. This approach generates a library of random combinatorial mutants limited to a precise region of human CYP1A1, namely amino acids 204-214 in which nine positions differ between CYP1A1 and CYP1A2. The resulting mutants present all combinations possible among these nine positions shifting mutated residues to their CYP1A2 counterpart. The mutants were cloned and expressed in an engineered Saccharomyces cerevisiae strain that has a microsomal oxido-reduction environment optimized for CYPs. This procedure resulted in yeast transformants that express a library of mutant CYP1A1. A subset of transformants were chosen at random, assayed for a typical CYP1A1 activity and the plasmidic DNA of functional clones was rescued and
A calendar is formed of a plurality of substrates. A first substrate carries indicia thereon which identifies selected time periods, such as days or months of the year. A second substrate is positioned adjacent to the first substrate. The second substrate defines a plurality of cavities dimensioned to individually retain a respective information carrying article, such as a web. Each of the cavities is corresponding supplied with a respective information carrying article. Each indicia on the first substrate is positionally associated with a respective cavity in the second substrate. A third substrate, positioned adjacent to the second substrate, is positioned to retain the information carrying articles releasably within the second substrate. The third substrate provides a rupturable cover over each of the cavities of the second substrate whereby upon the application of a sufficient lateral force on the information carrying article within a selected cavity, the article passes through the cover to a
Kyani offers three different products. Those products are Kyäni Sunrise, Kyäni Sunset, and Kyäni Nitro. http://ushap.kyaniviral.com/
Email: [email protected] Phone: (206) 667-5255. Currently, Dr. Chi is a staff scientist in the laboratory of Dr. Bruce Clurman in the Clinical Research Division of the Fred Hutch and is also affiliated with the laboratory of Dr. Robert Moritz at ISB. He has been developing proteomics-based tools to facilitate biological research. Specifically, he has pursued a proteomics approach to systematically identify the direct substrates of protein kinases. Mapping kinase and substrate relationships is critical for elucidating kinases functions and their signaling pathways. Despite the enormous interests and efforts in the field, it has remained a technical challenge, and the progress has been slow. He developed an in vitro-based method for proteome-wide identification of protein kinase substrates in cell lysates. This method utilized tools in biology, protein chemistry, and mass spectrometry and identified an unprecedented large number of candidate substrates for the human CDK2 kinase. Current in vitro ...
Email: [email protected] Phone: (206) 667-5255. Currently, Dr. Chi is a staff scientist in the laboratory of Dr. Bruce Clurman in the Clinical Research Division of the Fred Hutch and is also affiliated with the laboratory of Dr. Robert Moritz at ISB. He has been developing proteomics-based tools to facilitate biological research. Specifically, he has pursued a proteomics approach to systematically identify the direct substrates of protein kinases. Mapping kinase and substrate relationships is critical for elucidating kinases functions and their signaling pathways. Despite the enormous interests and efforts in the field, it has remained a technical challenge, and the progress has been slow. He developed an in vitro-based method for proteome-wide identification of protein kinase substrates in cell lysates. This method utilized tools in biology, protein chemistry, and mass spectrometry and identified an unprecedented large number of candidate substrates for the human CDK2 kinase. Current in vitro ...
Substrate identification needed? These standardized kinase substrate identification services are ideal for detection of substrates which might be phosphorylated by your protein kinases
Article{pmid25409537, Author=Thibodeaux, C. J. and Ha, T. and van der Donk, W. A. , Title={{A} price to pay for relaxed substrate specificity: a comparative kinetic analysis of the class {I}{I} lanthipeptide synthetases {P}roc{M} and {H}al{M}2}, Journal=J. Am. Chem. Soc., Year=2014, Volume=136, Number=50, Pages=17513--17529, Month=Dec ...
A technique for forming films of material (12) from a donor substrate (10). The technique has a step of introducing energetic particles (22) through a surface of a donor substrate (10) to a selected depth (20) underneath the surface, where the particles have a relatively high concentration to define donor substrate material (12) above the selected depth. Energy is provided to a selected region of the substrate to cleave a thin film of material from the donor substrate. Particles are introduced again into the donor substrate underneath a fresh surface of the donor substrate. A second thin film of material is then cleaved from the donor substrate.
Carbonyl reductase BaSDR1 has been identified as a potential ortho-haloacetophenone-specific biocatalyst for the synthesis of chiral 1-(2-halophenyl)ethanols due to its excellent stereoselectivity. However, the catalytic efficiency of BaSDR1 is far below the required level for practical applications. Thus, fine-tun
A module substrate consists of a substrate mounting electronic parts on one surface thereof, a conductor for electrically conducting the electronic parts mounted on the substrate to the other surface of the substrate, a conductive solder for attaching the conductor to a base substrate movably contacting the other surface of the substrate to electrically connect the electronic parts with the base substrate, and a deformable bushing for holding the conductor to maintain the attachment of the conductor to the base substrate regardless of whether the base substrate is moved.
Intelligent selections of enzyme substrates in microtiter plates for liquid phase assays (substrates for kinases, proteases and phosphatases).
Its not quite because silly a query as we may think. When you apply an anti wrinkle face cream, or any general skincare or anti aging product to a skin, one of the elements youll notice is that after youve rubbed it into the face it disappears. All of them, including the number one face…
A method for fabricating an LCD includes the steps of (a) loading a first substrate and a second substrate having seals formed thereon on a bonding chamber, (b) bonding the first and second substrates, (c) fixing the bonded first and second substrates, and (d) unloading the fixed first and second substrates.
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Enzyme- Enzymes are globular proteins, with a specific tertiary structure, which catalyse metabolic reactions in living organisms.. Enzymes are also known as biological catalysts because they speed up chemical reactions. They have a very specific 3D shape which is determined by their tertiary structure. The active site is the most important part of an enzyme. This is where the enzymes substrate binds. One theory of enzyme action is called the lock and key theory because the enzymes active site and the substrate are complementary in shape and charge.. Intacelluar Enzymes- Enzymes that catalyse reactions inside of cells. e.g Catalase, ATPsynthase, ATPase, DNA helicase, DNA polymerase, RNA polymerase, Lysosome hydrolytic enzymes. Extracellular Enzymes- Enzymes that catalyse reactions outside of cells.e.g digestive enymes. ...
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Press Release issued Jul 19, 2017: The biochemical reagents market is expanding at a rapid rate, due to use of these reagents in each and every section of health care and life sciences industries. A miscellaneous range of biochemical reagents is recognized for identification of specific enzymes in metabolism and for differentiation between bacteria and viruses. Classical biochemical tests are often used to identify microorganisms. Normally, the results are seen by change in color, formation of a specific product, or chemical reaction. In several cases, detection is based on the reaction of an enzyme with a certain substrate. Additionally, in order to detect specific enzymes or proteins by chemical reaction or complex building techniques are widely used, with the help of biochemical reagents. The end result leads to greater cognition of an unknown organism, protein, or assay.
Sigma-Aldrich provides many substrates to determine the activity of diverse enzymes. A wide range of fluorogenic and chromogenic substrates detect enzymatic activity optically.
Enzymes, commonly known as biocatalysts, are unique and highly specific globular proteins.. They accelerate chemical reactions without themselves undergoing any apparent change during the process.. They are produced within the cells but are capable of action outside the cells. The word enzyme was first introduced by Kuhne in 1878. Each enzyme usually acts on a single substrate and is said to be highly specific in its action.. According to lock and key hypothesis, the substrate molecules fit into the active sites located on the surface of the enzyme molecules just as one particu-lar key fits into one particular lock.. ...
This enzyme is synthesized as a proenzyme of 53 kDa that is converted to an active form of 22 kDa. cDNA sequences have been obtained for the mouse [3] and human [4] enzymes. In peptidase family M10 (interstitial collagenase family ...
The right price often makes the difference between a sale and a switch. In fact, across five different product or service attributes analyzed (price, service agreement, selection,...
Dr. Mc Allister responded: Debacterol. There are different products that can be used to help with the discomfort of canker sores. Everything from a |a href="/topics/topical-anesthetic" track_data="{
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TYPES OF ENZYME Oxidoreductases Transfer hydrogen and oxygen atoms or electrons from one substrate to another Transferases Transfer phosphate or methyl group from one substrate to another Hydrolases Hydrolysis a substrate Isomerases Change molecular form of the substrate Lyases Non-hydrolytic removal of a group or addition of a group to a substrate Ligases Join two […]. ...
Substrate, structural panel, bonded waterproofing The universal substrate for tiles Perfect covering No matter whether you work with mosaics or large format tiles, an absolutely level substrate with straight
A non competitive inhibitor is one in which doesnt compete with the substrate for the active site but binds away from it, this therefore changes the shape of the active site so denaturing it. This doesnt allow the enzyme substrate complex to form. ...
Chemicals for substrates available from Ladd Research online or call, (800) 451-3406, to order. Ladd Research has a large selection of chemicals needed to make substrates for sale.
A bottomless 6 channel slide used for cell culture applications with a self-adhesive underside to which own substrates can be ...
Hi folks, due to a technical issue at my host both of my forums are now lost to the void. Honestly, they had so little activity to begin with.
TY - JOUR. T1 - OXA-46, a new class D β-lactamase of narrow substrate specificity encoded by a blaVIM-1-containing integron from a Pseudomonas aeruginosa clinical isolate. AU - Giuliani, Francesco. AU - Docquier, Jean Denis. AU - Riccio, Maria Letizia. AU - Pagani, Laura. AU - Rossolini, Gian Maria. PY - 2005/5. Y1 - 2005/5. N2 - A novel OXA-type enzyme, named OXA-46, was found to be encoded by a gene cassette inserted into a class 1 integron from a multidrug-resistant Pseudomonas aeruginosa clinical isolate. The variable region of the integron also contained a blaVIM-1 metallo-β-lactamase cassette and a duplicated aacA4 aminoglycoside acetyltransferase cassette. OXA-46 belongs to the OXA-2 lineage of class D β-lactamases. It exhibits 78% sequence identity with OXA-2 and the highest similarity (around 92% identity) with another OXA-type enzyme detected in clinical isolates of Burkholderia cepacia and in unidentified bacteria from a wastewater plant. Expression of blaOXA-46 in Escherichia coli ...
What is enzyme substrate specificity? What are the importance of enzyme specificity? Classification of enzyme specificity, Different types of enzyme specificity: Bond specificity, Group specificity, Substrate specificity, Absolute Specificity, Optical or Stereo specificity, Geometrical specificity and Co-factor specificity. Learn more: Lecture Note in Specificity of Enzyme. You can DOWNLOAD the PPT by clicking on the download link below the preview…. ...
Use:. This mmp substrate can be used to assess activity of enzymes in the MMP family. The peptide sequence was described originally as a biosensor for MT1-MMP or MMP14 in "Simultaneous visualization of protumorigenic Src and MT1-MMP activities with fluorescence resonance energy transfer. Ouyang M, et al. Cancer Res. 2010 Mar 15;70(6):2204-12. doi: 10.1158/0008-5472.CAN-09-3698″. It demonstrates reasonably strong activity against MT1-MMP or MMP14 and MMP3, but has the highest activity against MMP9 with specificity constants, kcat/Km (M-1s-1), ranging from approximately 103 to 106. See also our Product Sheets for its substrate specificity profile. This substrate is not processed by ADAM family members. Typically, the peptide is dissolved in DMSO to make a stock solution of about 10mM concentration. When used for in vitro assays, the substrate is often used at about 10uM concentration. Remember to keep the DMSO concentration in the final reaction at 1% or below, to avoid DMSO effects on the ...
Novel glycine oxidase (GlyOX) from Marinomonas mediterranea depends on cysteine tryptophilquinone (CTQ) and catalyzes the oxidative deamination of glycine to produce a glyoxylate, ammonia, and hydrogen peroxide. M. mediterranea GlyOX genes (goxA and goxB) were cloned and recombinant GlyOX was heterologously expressed by E. coli. The purification of recombinant GlyOX was carried out by metal affinity and DEAE-Toyopearl 650M column chromatographies. M. mediterranea GlyOX was homotetramic with a molecular mass of 76kDa and showed optimum activity around 30°C and at pH 5.0, and stability below 50°C and between pH 5.0 to 9.0. M. mediterranea GlyOX shows a strict substrate specificity toward glycine, and the Michaelis constant for glycine was 0.5mM. M. mediterranea GlyOX could determine the quantity of glycine in human serum and human blood plasma with high sensitivity. This study revealed the catalytic and structural properties of M. mediterranea GlyOX with high substrate specificity. ...
Background: Our understanding of how fungi evolved to develop a variety of ecological niches, is limited but of fundamental biological importance. Specifically, the evolution of enzymes affects how well species can adapt to new environmental conditions. Feruloyl esterases (FAEs) are enzymes able to hydrolyze the ester bonds linking ferulic acid to plant cell wall polysaccharides. The diversity of substrate specificities found in the FAE family shows that this family is old enough to have experienced the emergence and loss of many activities. Methodology/Principal Findings: In this study we evaluate the relative activity of FAEs against a variety of model substrates as a novel predictive tool for Ascomycota taxonomic classification. Our approach consists of two analytical steps; (1) an initial unsupervised analysis to cluster the FAEs substrate specificity data which were generated by cultivation of 34 Ascomycota strains and then an analysis of the produced enzyme cocktail against 10 substituted
AmpC BER is an extended substrate spectrum class C beta-lactamase with a two-amino-acid insertion in the R2 loop compared with AmpC EC2. The crystal structures of AmpC BER (S64A mutant) and AmpC EC2 were determined. Structural comparison of the two proteins revealed that the insertion increases the conformational flexibility of the R2 loop. Two citrate molecules originating from the crystallization solution were observed in the active site of the S64A mutant. One citrate molecule makes extensive interactions with active-site residues that are highly conserved among class C beta-lactamases, whereas the other one is weakly bound. Based on this structural observation, it is demonstrated that citrate, a primary metabolite that is widely used as a food additive, is a competitive inhibitor of two class C beta-lactamases (AmpC BER and CMY-10). Consequently, the data indicate enhancement of the flexibility of the R2 loop as an operative strategy for molecular evolution of extended-spectrum class C ...
Vanillyl alcohol oxidase (VAO) and eugenol oxidase (EUGO) are flavin-dependent enzymes that catalyse the oxidation of para-substituted phenols. This makes them potentially interesting biocatalysts for the conversion of lignin-derived aromatic monomers to value-added compounds. To facilitate their biocatalytic exploitation, it is important to develop methods by which variants of the enzymes can be rapidly screened for increased activity towards substrates of interest. Here, we present the development of a screening assay for the substrate specificity of para-phenol oxidases based on the detection of hydrogen peroxide using the ferric-xylenol orange complex method. The assay was used to screen the activity of VAO and EUGO towards a set of twenty-four potential substrates. This led to the identification of 4-cyclopentylphenol as a new substrate of VAO and EUGO and 4-cyclohexylphenol as a new substrate of VAO. Screening of a small library of VAO and EUGO active-site variants for alterations in their
Phytaspase is a member of the plant subtilisin-like protease family, and is commonly distinguished from the other members by its unusual and extremely high specificity towards its substrates, which resembles that of the animal caspases. Similarly to the animal caspases, the phytaspase is a cell death promoting protease. The name phytaspase comes from phyto- (lat. for plant) and -aspase (aspartate-directed protease), similarly to caspases. The phytaspase displays a strict substrate specificity, which resembles that of the animal caspase-3. It recognizes a tetrapetide motive within a target protein and introduces a peptide bond break following an aspartate residue, which is crucial for the hydrolysis. Theoretical speculations, based on a 3D model predictions have been made, pointing to the histidine 331 of the phytaspase peptide chain, that might interact with the Asp in the target peptide and thereby guide the recognition. The phytaspase displays a structure, common to the subtilisin-like ...
0078] The coating composition according to the instant invention may be applied to a substrate. Exemplary suitable substrates include, but are not limited to, sheet, non-woven material, woven material, film, foams, and the like. Such substrate may comprise organic based materials, inorganic materials, and combinations thereof. The substrate may, for example, comprise a cellulose based material, a natural polymeric based material, a synthetic polymeric based material, a metal based material, a mineral based, and combinations thereof. The substrate may be porous, for example, micro-porous. The coating composition may be applied to the substrate via a conventional method for applying a coating composition. Such methods are generally known, and include, but are not limited to spraying, dipping, roll coating, blade coating, curtain coating, printing techniques such as flexography and rotogravure, size press, metered size press, screen coating, rod coating combinations thereof, and the like. The ...
Galactokinase; Sugar-1-kinase with a strict substrate specificity for the alpha-anomeric configuration of D-galacturonic acid (D-GalA) and ATP. Involved in the biosynthesis of UDP-galacturonic acid (UDP-GalA) from the salvaged GalA that is released during growth- dependent cell wall restructuring (424 aa ...
SandeepWeb is a blog for research and reviews of different products that will help users to decide if the product is best for them or not.
SandeepWeb is a blog for research and reviews of different products that will help users to decide if the product is best for them or not.
Caspase-3 is a cysteine protease that hydrolyzes diverse intracellular proteins during programmed cell death (known as apoptosis). It has been a popular target for drug design against abnormal cell death for more than a decade. No approved caspase based drug, however, is available so far. Therefore, structural insights about the substrate recognition of caspase-3 are needed for the future development of caspase-3 based inhibitors and drugs. In this study, crystal structures of recombinant caspase-3 in complex with seven substrate analog inhibitors, including acetyl (Ac)-DEVD-aldehyde (Cho), Ac-DMQD-Cho, Ac-IEPD-Cho, Ac-YVAD-Cho, Ac-WEHD-Cho, Ac-VDVAD-Cho, and tert-butoxycarbonyl (Boc)-D-fluoromethylketone (Fmk), have been analyzed in combination with enzyme kinetic data and computational models. Seven crystal structures were determined at resolutions of 1.7-2.6Å. The binding conformation of each inhibitor residue at P1-P4 position was analyzed. The negative P1 aspartic acid side chain is exclusively
Identification of Crucial Amino Acids in Mouse Aldehyde Oxidase 3 That Determine Substrate Specificity. . Biblioteca virtual para leer y descargar libros, documentos, trabajos y tesis universitarias en PDF. Material universiario, documentación y tareas realizadas por universitarios en nuestra biblioteca. Para descargar gratis y para leer online.
We are interested in the mechanistic and molecular relationships between catalytic activity, conformational changes and microenvironment of ABC transporters. P-glycoprotein (ABCB1, Pgp) is in the focus of our interest; we have currently extended our work to ABCG2 (BCRP) and plan to do similar studies on MRP1 (ABCC1). The members of the ABC superfamily of membrane transporters are involved in the regulation of the uptake into and distribution within our body of physiological substrates as well as various xenobiotics, drugs. Due to their wide substrate spectrum, a consequence of their preference for lipophylic compounds, they also play a critical role in the multidrug resistance phenomenon severely limiting therapeutical success in cancer. Our ambition is to understand the molecular details of their catalytic cycle and the intimate molecular interactions with their microenvironment, as well as to apply the knowledge obtained at the cell/molecule level in the context of the whole organism, in ...
Substrate specificity of hyaluronidases tested on polyacrylamide gel with incorporated chondroitin sulfate.Protein content per 2 µl dot is indicated in bracket
Has an unusual substrate specificity for synthetic organophosphate triesters and phosphorofluoridates. All of the phosphate triesters found to be substrates are synthetic compounds. The identity of any naturally occurring substrate for the enzyme is unknown. Has no detectable activity with phosphate monoesters or diesters and no activity as an esterase or protease. It catalyzes the hydrolysis of the insecticide paraoxon at a rate approaching the diffusion limit and thus appears to be optimally evolved for utilizing this synthetic substrate ...
The primary specificity residue of a substrate or an inhibitor, called the P1 residue, is responsible for the proper recognition by the cognate enzyme. This residue enters the S1 pocket of the enzyme and establishes contacts (up to 50%) inside the proteinase substrate cavity, strongly affecting its specificity. To analyze the influence on bovine α-chymotrypsin substrate activity, aromatic non-proteinogenic amino acid residues in position P1 with the sequence Ac-Phe-Ala-Thr-XAnb 5,2-NH2 were introduced: L-pyridyl alanine (Pal), 4-nitrophenylalanine - Phe(p-NO2), 4-aminophenylalanine - Phe(p- NH2), 4-carboxyphenylalanine Phe(p-COOH), 4-guanidine phenylalanine - Phe(p-guanidine), 4-methyloxycarbonylphenylalanine - Phe(p-COOMe), 4-cyanophenylalanine - Phe(p-CN), Phe, Tyr. The effect of the additional substituent at the phenyl ring of the Phe residue was investigated. All peptides contained an amide of 5-amino-2-nitrobenzoic acid, which served as a chromophore. Kinetic parameters (kcat, KM and ...
Many predicted (phospho)lipases are poorly characterized with regard to their substrate specificities and physiological functions. Here ...
Motivation:In silico methods are being widely used for identifying substrates for various kinases and deciphering cell signaling networks. However, most of the available phosphorylation site prediction methods use motifs or profiles derived from a known data set of kinase substrates and hence, their applicability is limited to only those kinase families for which experimental substrate data is available. This prompted us to develop a novel multi-scale structure-based approach which does not require training using experimental substrate data.. Results:In this work, for the first time, we have used residue-based statistical pair potentials for scoring the binding energy of various substrate peptides in complex with kinases. Extensive benchmarking on Phospho.ELM data set indicate that our method outperforms other structure-based methods and has a prediction accuracy comparable to available sequence-based methods. We also demonstrate that the rank of the true substrate can be further improved, if ...
The RAS/MAPK pathway has been intensively studied [1-4], with constitutive activation of ERK1 and ERK2 found frequently in human cancer cells from a variety of tissues (e.g., lung, pancreas, colon, ovary, kidney, skin, and thyroid) [13]. Amplification, overexpression, or mutations in RTKs and genetic alterations in upstream components of the MAPK pathway, including KRAS, NRAS, HRAS, CRAF, BRAF, MEK1, and MEK2, alter cell signaling in tumors. In clinical practice and clinical trials, small molecules targeting RTKs or components in the MAPK cascade are used to treat cancer [1, 3, 4]. MEK1 and MEK2 are ideal targets; not only do they play a key role in tumor development and progression [3, 4], they have narrow substrate specificities and distinctive structural characteristics.. MEK activation through the MAPK signaling cascade is necessary for mammalian cell transformation, and constitutively active MEK mutants promote transformation of fibroblast cells [14, 15]. Furthermore, MEK inhibitors inhibit ...
within the GH-J clan. Moreover, besides the effect of substrate entrance on its own, we strongly suggest that a highly conserved arginine residue (in the RDP motif) rather than the previously proposed Tyr motif (not conserved) provides the proton to increase the pKa of the acid-base catalyst ...
Chaetoviridins constitute a large family of structurally related secondary metabolites isolated from Chaetomium fungi. To elucidate the biosynthesis pathway and understand how the chemical diversity of chaetoviridins is generated, gene deletion and in vitro characterization of the four post-PKS modifications enzymes were undertaken. CazL and CazP were identified to have substrate promiscuity that facilitates the formation of nonchlorinated analogues. In addition, enzymatic oxidation and reduction combined with spontaneous dehydration and lactonization of the intermediates further expand the chemical diversity ...
Thus, when a great deal of substrate is altered by an enzyme every minute, the reaction is said to be proceeding at a rapid rate.. In enzyme reaction rates, the rate depends on the CONCENTRATION of the enzyme and the CONCENTRATION of the substrate (CONCENTRATION rather than AMOUNT). Concentration refers to amount in a given volume of solution. As previously mentioned, it has been calculated that enzyme mediated reactions occur 1 x 109 times faster than the same reactions without enzymes.. In most enzyme reactions, enzyme concentration is small compared to the substrate concentration. Therefore, the rate of the reaction becomes proportional to the concentration of the enzyme. If the enzyme concentration is doubled, the reaction rate is doubled. At low substrate concentrations, the rate of the reaction is proportional to the substrate concentration, but at higher substrate concentrations the reaction rate is independent of substrate concentration. That is, further increase in the amount of ...
An apparatus includes a first substrate; and a second substrate coupled to the first substrate, characterized in that, to control formation of a segregated phase domain structure within a chemical reaction product by controlling an amount of a constituent of a precursor that is present per unit surface area, at least one member selected from the group consisting of the first substrate and the second substrate defines a substantially regularly periodically varying relief with respect to basal spatial location.
PubMed comprises more than 30 million citations for biomedical literature from MEDLINE, life science journals, and online books. Citations may include links to full-text content from PubMed Central and publisher web sites.
BioAssay record AID 1078796 submitted by ChEMBL: Inhibition of human FER catalytic domain expressed in baculovirus assessed as substrate phosphorylation using fluorescence-labelled peptides as substrate at 9 uM after 90 mins by microfluidic peptide phosphorylation assay.
BioAssay record AID 1078965 submitted by ChEMBL: Inhibition of human FGFR4 catalytic domain expressed in baculovirus assessed as substrate phosphorylation using fluorescence-labelled peptides as substrate at 0.06 uM after 90 mins by microfluidic peptide phosphorylation assay.
Identifying the substrates of protein kinases to understand their modes of action has been undertaken by various approaches and remains an ongoing challenge
1UN4: Crystallographic Studies on Structural Features that Determine the Enzymatic Specificity and Potency of Human Angiogenin: Thr44, Thr80 and Residues 38-41
At the atomic scale, we are interested in providing detailed three-dimensional information about the nature of complex metallocofactors to help understand how protein environment modulates reactivity. At the protein scale, we are interested in seeing how enzymes are constructed to control substrate access and specificity, and how they prevent loss of reactive intermediates or damage to expensive cofactors. At the largest scale, that of protein complexes, we want to know how proteins interact and how those interactions explain the observed behavior. Protein complexes are often large, have multiple distinct states, and can have large inter- and intrasubunit motions; therefore a single "snapshot" usually does not tell the entire story.. ...
Negative cooperativity of ATP and substrate binding and positive cooperativity of ADP and substrate binding.(a) The effect of ATP concentration on substrate Km
Shop Multisubstrate adapter protein ELISA Kit, Recombinant Protein and Multisubstrate adapter protein Antibody at MyBioSource. Custom ELISA Kit, Recombinant Protein and Antibody are available.
You see the word more and more, but what does it say about what it's on? Here are tips for fruits and vegetables, dairy and meat, cosmetics, processed foods and cotton and coffee.
antibody-antibodies.com is the marketplace for research antibodies. Find the right antibody for your research needs. Cleavage specificity of cucumisin, a plant serine protease.
techreport{5f99741f-59be-441f-9f2c-c234b4626641, abstract = {In this paper a method for the analysis of a frequency selective surface (FSS),br/,,br, supported by a bianisotropic substrate is presented. The frequency selective,br/,,br, structure is a thin metallic pattern - the actual FSS - on a plane supporting,br/,,br, substrate. Integral representations of the fields in combination with the,br/,,br, method of moments carried out in the spatial Fourier domain are shown to,br/,,br, be a fruitful way of analyzing the problem with a complex substrate. This,br/,,br, approach results in a very general formulation in which the supporting substrate,br/,,br, can have arbitrary bianisotropic properties. The bianisotropic slab,br/,,br, can be homogeneous, stratified, or it can have continuously varying material,br/,,br, parameter as a function of depth. The analysis presented in this paper is,br/,,br, illustrated in a series of numerical examples. Results for isotropic, anisotropic,br/,,br, and ...
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A method of performing treatment under a reduced pressure for processing a substrate placed in a chamber, includes the steps of providing a heater within the chamber, for heating the substrate, placing the substrate on a susceptor, the substrate being placed above the heater within the chamber, chucking the substrate on the susceptor above the heater, heating the substrate with the heater, and evacuating the interior of the chamber to provide a reduced pressure environment.
A process of making a multilayer printed wiring board assembly. The process includes the steps of providing a first and a second substrate made of a dielectric material; depositing a first wiring pattern on the first substrate and a second wiring pattern on the second substrate with a conductive material; depositing a dielectric material on the first and second wiring patterns and defining a via connecting zone on the first and the second wiring pattern for communicating signals between the first and the second wiring pattern by exposing a selective portion of the first and second wiring patterns; depositing a conductive bonding material on the via connecting zone of one of the first and the second wiring pattern; arranging the first and the second substrate in sandwiched juxtaposition such that the via connecting zones of the first and the second wiring pattern are opposite each other and in substantial alignment with each other so that the conductive bonding material deposited on the one of the via
A komplement rendszer aktiválódásának lektin útja az egyik első védelmi vonalnak tekinthető a szervezet fertőzések elleni védekezésében. A mannóz kötő lektin (MBL) baktérium felszínhez való kötődése után szerin proteáz zimogének (MASP= MBL-kötött szerin proteáz) aktiválódnak, melyek többféle mechanizmus révén járulnak hozzá az idegen mikroorganizmus megsemmisítéséhez ill. eltávolításához. Munkánk során felderítettük, a proteolitikus kaszkádrendszer beindításáért felelős MASP-2 enzim autoaktiválódásásnak mechanizmusát atomi szinten. Felfedeztük a MASP-2 egy eddig ismeretlen biológiai funkcióját, amely kapcsolatot teremt a véralvadási és a komplement kaszkád között. A MASP-2 hasítja és aktiválja a protrombint. Ugyancsak részletesen tanulmányoztuk a MASP-1 trombin-szerű aktivitását is. Ezek az eredmények arra utalnak, hogy a vérben lévő két proteolitikus kaszkádrendszer szoros evolúciós és funkcionális ...
A method for bonding together two or more acid-doped polybenzimidazole films is provided. The method includes, in the following order: placing a first acid-doped polybenzimidazole film on a first substrate to form a first film/ substrate assembly and placing a second acid-doped polybenzimidazole film on a second substrate to form a second film/substrate assembly; heating the first and second film/substrate assemblies to a temperature sufficient to soften the first and second acid-doped polybenzimidazole films; positioning the second film/substrate assembly atop the first film/substrate assembly, such that the first acid-doped polybenzimidazole film is in contact with the second acid-doped polybenzimidazole film and such that polybenzimidazole polymer chains of the first acid-doped polybenzimidazole film interact with polybenzimidazole polymer chains of the second acid-doped polybenzimidazole film; and re- hydrolyzing the first and second acid-doped polybenzimidazole films, such that the
No binding heat in Substrate and Enzyme ITC - posted in Molecular Biology: Dear all, I did the ITC (isothermal titration calorimetry) between an enzyme with its known substrate on Microcal ITC200. The concentartion of the ligand (substarte) is 1mM; and the concentration of the enzyme is 100 uM; 1.8 ul/injection X 22 injection; However, there is no heat of binding observed in the raw data (isotherms). (dilution of substrate(substrate to buffer control) seems to consume some heat; dilution o...
Mapping kinase‐substrate interactions demands robust methods to rapidly and unequivocally identify substrates from complex protein mixtures
The types of specificity that can be ascribed to lipases include 1. Substrate specificity The enzyme shows a different rate of lipolysis of various
iHOP - Information Hyperlinked over Proteins. iHOP provides the network of genes and proteins as a natural way of accessing the millions of abstracts in PubMed. By employing genes and proteins as hyperlinks between sentences and abstracts, the information in PubMed becomes bound together into one navigable resource. A Gene Network for Navigating the Literature, Nature Genetics 36, 664 (2004). www.ihop-net.org/UniPub/iHOP/
3CMS: Engineering enzyme subsite specificity: preparation, kinetic characterization, and X-ray analysis at 2.0-A resolution of Val111Phe site-mutated calf chymosin.
Published in J. Phys. Chem. B, 2010. Recommended citation: Jing Zhao, Chang Lu, Stefan Franzen*, J. Phys. Chem. B, 2015,119 (40), pp 12828-12837. http://pubs.acs.org/doi/abs/10.1021/acs.jpcb.5b07126 ...
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I dont think theres ever been a resistance to the idea of unstructured sections, since any work I did examining protein structures would show me that the crystals were missing pieces, particularly at both end of the protein, because those were unstructured even in the crystals. Often also loops around binding sites. So when I chatted with Keith I thought he was exaggerating a bit about resistance to the idea. I told him so, then he agreed that people with experience in structure were more open to these ideas, but not that unstructured stuff could improve specificity. I disagree with improved specificity (I told him so too), but I think that might be quite hard to test. For that we had mostly anecdotal examples, rather than real numbers in calories and all comparing unstructured to structured and highly discriminating proteins. But I have not followed the area too much since I moved into genomics. Whats your take? Is there better demonstration of this specificity claim?. Delete ...
See underneath concerning why we think glucose is not the most effective substrate to diagnose SIBO. Working with glucose only given that the test substrate can pass up a lot of SIBO beneficial situations.To prepare for your test, its a good idea to request your health care provider about which test you can be taking, and also to ask about any die. ...
Why does increasing substrate concentration increased rate of reaction, An explanation of the effect of substrate concentration, temperature and pH on. If you plotted a graph of initial reaction rate against the concentration of a. Increasing the concentration any more makes no difference to the rate of the reaction.
I have been contemplating what substrate I will use when I finally grow some cubes (hopefully sometime, though I live in CA now so itll make the task a bit harder w/ shipping issues). I remember
A lithographic apparatus is disclosed that has a first substrate table arranged to hold a substrate and a second substrate table arranged to hold a substrate, an imprint template holder arranged to ho
Dermoshop Oy is developing and marketing skin care and makeup products under the brand name Dermosil. All the products in the Dermosil product assortment are made of high quality ingredients and are subjected to a thorough dermatological testing process. The product assortment includes basic skin care products for the entire family, highly developed makeup products and trendy special care products. The product assortment contains about 400 different products.. ...
Dermoshop Oy is developing and marketing skin care and makeup products under the brand name Dermosil. All the products in the Dermosil product assortment are made of high quality ingredients and are subjected to a thorough dermatological testing process. The product assortment includes basic skin care products for the entire family, highly developed makeup products and trendy special care products. The product assortment contains about 400 different products.. ...
The selective chemical transformation of cellulose into different products by hydrothermal treatment under controlled conditions has been achieved in the labs
Melet, A., Marques-Soares, C., Schoch, G. A., Macherey, A. C., Jaouen, M., Dansette, P. M., Sari, M. A., Johnson, E. F., Mansuy, D. Analysis of human cytochrome P4502C8 substrate specificity using a substrate pharmacophore and site-directed mutants Biochemistry 2004 43:15379-15392 DOI:10.1021/bi0489309 PMID:15581350 ...
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The substrate specificity is very similar to that of EC 2.3.1.179 with the exception that the latter enzyme is far more active with palmitoleoyl-ACP (C(16)-Delta(9)) as substrate, allowing the organism to regulate its fatty-acid composition with changes in temperature ...
Aniline dioxygenase is a multicomponent Rieske nonheme-iron dioxygenase enzyme isolated from Acinetobacter sp. strain YAA. Saturation mutagenesis of the substrate-binding pocket residues, which were identified using a homology model of the α subunit of the terminal dioxygenase (AtdA3), was used to probe the molecular determinants of AtdA substrate specificity. The V205A mutation widened the substrate specificity of aniline dioxygenase to include 2-isopropylaniline, for which the wild-type enzyme has no activity. The V205A mutation also made 2-isopropylaniline a better substrate for the enzyme than 2,4-dimethylaniline, a native substrate of the wild-type enzyme. The I248L mutation improved the activity of aniline dioxygenase against aniline and 2,4-dimethylaniline approximately 1.7-fold and 2.1-fold, respectively. Thus, it is shown that the α subunit of the terminal dioxygenase indeed plays a part in the substrate specificity as well as the activity of aniline dioxygenase. Interestingly, the ...
Insulin receptor substrates (IRS)-5 and -6 are two recently identified members of the IRS family. We investigated their roles as insulin receptor substrates and compared them with Src-homology-2-containing (Shc) protein, a well-established substrate. Bioluminescence resonance energy transfer (BRET) experiments showed no interaction between the receptor and IRS-5, while interaction with IRS-6 was not enhanced by insulin. By contrast, Shc showed an insulin-induced BRET response, as did a truncated form of IRS-1 (1-262). While Shc was heavily phosphorylated after stimulation of the insulin receptor, IRS-5 and -6 showed very weak phosphorylation levels. These results suggest that, although these two adaptors have previously been proposed as substrates for the insulin receptor, they are poor substrates for the insulin receptor. This calls into question their relevance to insulin signalling.
The substrate recognition mechanisms in chaperonins Chaperonins are a family of proteins devoted to assisting the folding of other proteins. They are large oligomers assembled into ring structures that enclose a cavity in which folding takes place. For this process to occur, the chaperonin must first recognize and interact with the unfolded polypeptide, then undergo a conformational change upon nucleotide binding that results in the closure of the cavity which in turn mediates the folding reaction inside the cavity. Although this general mechanism seems to apply to every chaperonin studied so far, there exist two different modes of interaction between the chaperonin and the substrate. The first occurs mainly through the interaction between the exposed hydrophobic residues of the unfolded polypeptides and those of the chaperonin substrate binding site, as elucidated for the chaperonin GroEL from E. coli. The second type of mechanism has been described so far only for the cytosolic chaperonin CCT ...
Papain-like cysteine proteases are important for the survival of the flagellated protozoa Trypanosoma cruzi, the causative agent of Chagas Disease. the lysosomal cysteine protease designated as cruzipain or cruzain, is the archetype of a multigene family of related isoforms. We investigated the substrate specificity of the cruzipain 2 isoform using internally quenched fluorogenic substrates. We found that cruzipain 2 and cruzain differ substantially regarding the specificity in the S-2, S-1() and S-2() pockets. Our study indicates that cruzipain 2 has a more restricted specificity than cruzain, suggesting that these isoforms might act on distinct natural substrates ...
Deoxyribonucleoside kinases catalyze the phosphorylation of deoxyribonucleosides to the corresponding deoxyribonucleoside monophosphates (dNMPs). They are the key enzymes in the salvage of deoxyribonucleosides originating from extra‐ or intracellular breakdown of DNA. Subsequently, dNMPs are phosphorylated into diphosphates (dNDPs) and triphosphates (dNTPs), which are the precursors of DNA. Deoxyribonucleoside kinases play a key role in the chemotherapeutic treatment of cancer and viral diseases, as they catalyze the first, and often rate‐limiting step of nucleoside analog activation by phosphorylation (Arnér and Eriksson, 1995). Native and genetically engineered deoxyribonucleoside kinases from different organisms are also attractive candidates for use in cancer gene therapy as suicide enzymes (Christians et al., 1999; Kokoris et al., 1999; Knecht et al., 2000a; Zheng et al., 2000). The basic concept here is to transduce cancer or virus‐infected cells with a gene encoding a ...
Glycosidases hydrolyse the glycosidic bond in carbohydrates. Structural studies of three glycosidases with different substrate specificities are presented in this work.. Dextranase catalyzes the hydrolysis of α-1,6-glycosidic linkage in dextran polymers. The structure of dextranase, Dex49A, from Penicillium minioluteum was solved in the apo-enzyme (1.8 Å resolution) and product-bound (1.65 Å resolution) forms. The main domain of the enzyme is a right-handed β-helix, which is connected to a β-sandwich domain at the N-terminus. Using NMR spectroscopy the reaction course was shown to occur with net inversion at the anomeric carbon. A new clan is suggested that links glycoside hydrolase (GH) families 28 and 49.. Endo-β-1,4-D-mannanase catalyzes the depolymerization of β-1,4-mannan polymers. The structure of endo-1,4-β-mannanase Man5A from blue mussel Mytilus edulis has been determined at 1.6 Å resolution. Kinetic analysis of Man5A revealed that the enzyme requires at least 6 subsites for ...
The invention provides a method for determining an amino acid sequence motif for a phosphorylation site of a protein kinase. In the method of the invention, a protein kinase is contacted with an oriented degenerate peptide library, peptides within the library which are substrates for the kinase are converted to phosphopeptides and the phosphopeptides are separated from non-phosphorylated peptides. The isolated phosphopeptides are sequenced and an amino acid sequence motif for the phosphorylation site is determined based upon the relative abundance of different amino acids residues at each degenerate position. The invention also provides peptide substrates for protein kinase A, cell cycle control kinases, src family kinases, the EGF receptor and p92.sup.c-fps/fes based upon amino acid sequence motifs for the phosphorylation sites of these kinases.
Glycoside hydrolases (O-Glycosyl hydrolases) EC 3.2.1. are a widespread group of enzymes that hydrolyse the glycosidic bond between two or more carbohydrates, or between a carbohydrate and a non-carbohydrate moiety. A classification system for glycosyl hydrolases, based on sequence similarity, has led to the definition of numerous different families. This classification is available on the CAZy (CArbohydrate-Active EnZymes) web site. Because the fold of proteins is better conserved than their sequences, some of the families can be grouped in clans. As of October 2011, CAZy includes 128 families of glycosyl hydrolases and 14 clans. Glycoside hydrolase family 1 Glycoside hydrolase family 2 Glycoside hydrolase family 3 Glycoside hydrolase family 4 Glycoside hydrolase family 5 Glycoside hydrolase family 6 Glycoside hydrolase family 7 Glycoside hydrolase family 8 Glycoside hydrolase family 9 Glycoside hydrolase family 10 Glycoside hydrolase family 11 Glycoside hydrolase family 12 Glycoside ...
Substrate binding and specificity[edit]. The structures of several HAT domains bound to acetyl-CoA and histone substrate ... 4 Substrate binding and specificity *4.1 Lysine selectivity *4.1.1 GNAT family ... The formation of multisubunit complexes has been observed to modulate the substrate specificity of HATs.[10] In general, while ... For example, the lysine specificity of MYST family HATs toward their histone substrates becomes more restricted when they ...
Implications for substrate specificity". J. Biol. Chem. 273 (34): 21714-20. doi:10.1074/jbc.273.34.21714. PMID 9705307.. ...
Use of Oriented Peptide Libraries to determine phosphopeptide binding specificity and protein kinase substrate specificity[edit ... 2.2 Use of Oriented Peptide Libraries to determine phosphopeptide binding specificity and protein kinase substrate specificity ... This approach was used to characterize the substrate specificity of a large number of protein kinases. The kinase specificity ... P as a substrate, in fact required PtdIns(5)P as a substrate to produce PtdIns(4,5)P2.[34] Further research demonstrated that ...
Retrieved from Encyclopædia Britannica Online: [1] Miller FD, Chapman JL, Queener SW (1992). "Substrate specificity of ... This allows for the binding of the substrate ACV to the deprotonated thiol group of the cysteine residue. This ligation of the ... and two water molecules in the absence of a bound substrate. Just two histidine residues and one aspartic acid residue are ... "Structure of isopenicillin N synthase complexed with substrate and the mechanism of penicillin formation". Nature. 387 (6635): ...
These enzymes halogenate without substrate specificity and regioselectivity. The first tryptophan 7-halogenase was isolated in ... These results suggest that for larger substrates, perhaps the entirety of the substrate molecule is not enclosed in the active ... Enzyme activity was evolved with a known substrate that bears structural similarity to the target substrate, then variants with ... Site-directed mutagenesis has also been employed for expanding substrate scope. Tryptophan substrate interacts with numerous ...
It is now known that isomerases have broad substrate specificity. Most pentoses and some hexoses are all substrates for D- ... Metal site 1 binds the substrate tightly, while metal site two binds the substrate loosely. Both share an acid residue Glutamic ... The first metal, mentioned earlier, coordinates to O3 and O4, and is used to dock the substrate. In the isomerization of xylose ... state consists of a high energy carbonium ion that is stabilized through all the metal interactions with the sugar substrate. ...
Substrate specificities. Preferred cofactor. Catalytic preference. Tissue distribution. Expression profile. Pathology 1. ... HSD17B10: Also known as 2-methyl-3-hydroxybutyryl-CoA dehydrogenase (MHBD). Substrates include steroids, neurosteroids, fatty ...
A second precaution with respect to predicting FMO enzyme substrate specificity is that factors other than size and charge must ... substrate specificity and role in drug metabolism". Curr. Drug Metab. 1 (2): 181-191. doi:10.2174/1389200003339135. PMID ... This enzyme has a wide substrate specificity, including the dietary-derived tertiary amines trimethylamine, tyramine and ... Other drug substrates have been used for both in vitro and in vivo analyses. ... FMO3 is the most abundantly expressed FMO in ...
... comparison of its substrate specificity with that of other SAP kinases". The EMBO Journal. 16 (12): 3563-71. doi:10.1093/emboj/ ... Dual specificity mitogen-activated protein kinase kinase 6 also known as MAP kinase kinase 6 (MAPKK 6) or MAPK/ERK kinase 6 is ... MAPKK 6 is a member of the dual specificity protein kinase family, which functions as a mitogen-activated protein (MAP) kinase ... "Nuclear export of the stress-activated protein kinase p38 mediated by its substrate MAPKAP kinase-2". Current Biology. 8 (19): ...
"Dynamin is a minibrain kinase/dual specificity Yak1-related kinase 1A substrate". The Journal of Biological Chemistry. 277 (20 ... "Specificity of the binding of synapsin I to Src homology 3 domains". The Journal of Biological Chemistry. 275 (38): 29857-67. ... bind to the proline-rich region of synaptojanin 1 at distinct sites that display an unconventional binding specificity". The ...
Villiers BR, Hollfelder F (March 2009). "Mapping the limits of substrate specificity of the adenylation domain of TycA". ... whereas the ability to act upon different substrates is called substrate promiscuity or substrate ambiguity. The term latent ... Enzymes are evolved to catalyse a particular reaction on a particular substrate with a high catalytic efficiency (kcat/KM, cf. ... When the specificity of enzyme was probed, it was found that it was highly selective against natural amino acids that were not ...
In addition to controlling activity, the allosteric mechanism also regulates the substrate specificity and ensures the enzyme ... RNR1 contains both allosteric sites, mediating regulation of substrate specificity and activity. Depending on the allosteric ... Reduction of NDP substrates occurs under aerobic conditions. Class I reductases are divided into IA and IB due to differences ... The substrates for RNR are ADP, GDP, CDP and UDP. dTDP (deoxythymidine diphosphate) is synthesized by another enzyme ( ...
Implications for substrate specificity". J. Biol. Chem. 273 (34): 21714-20. doi:10.1074/jbc.273.34.21714. PMID 9705307. ... and is also found in a variety of proteins with phosphorylated substrates. These include ATP synthase (α and β subunits), ...
Multiplicity and substrate specificity". Biochimica et Biophysica Acta. 657 (2): 457-67. doi:10.1016/0005-2744(81)90331-4. PMID ... Tachibana Y, Yamashita K, Kobata A (1982). "Substrate specificity of mammalian endo-beta-N-acetylglucosaminidase: study with ... PNGase F lacks selectivity for outer carbohydrate structure, resulting in broad specificity, making it a useful tool for ... These endoglycosidases have more specificity in cleavage and are less sensitive to protein conformation than PNGase F. All of ...
Substrate and inhibitor specificity. J Biol Chem 1979, 254: 2346-2352. Cass CE, Selner M, Phillips JR: Resistance to 9-beta-D- ... Compounds that are substrates for AdK include the N-nucleosides toyocamycin, tubercidin and 6-methylmecaptopurine riboside; the ... identification of a novel motif implicated in phosphate and magnesium ion binding and substrate inhibition. Biochemistry 2002, ...
Substrate and cofactor specificity". J. Biol. Chem. 234 (8): 2129-32. PMID 13673025. Moorefield HH (1956). "Purification of DDT ... this enzyme has one substrate, 1,1,1-trichloro-2,2-bis(4-chlorophenyl)ethane, and two products, 1,1-dichloro-2,2-bis(4- ...
Specificity[edit]. Proteolysis can be highly promiscuous such that a wide range of protein substrates are hydrolysed. This is ... Promiscuous proteases typically bind to a single amino acid on the substrate and so only have specificity for that residue. For ... TopFIND - database of protease specificity, substrates, products and inhibitors. *MEROPS - Database of protease evolutionary ... have high specificity and only cleave very restricted set of substrate sequences. They are therefore a common target for ...
However, differences exist which unsurprisingly account for differences in substrate specificity among peroxidases. This ... implicating this residue or its peculiar functional group in substrate specificity. Cyanide binds very tightly to mammalian ... The distal face is used for substrate binding and catalysis. The crystal structures of MPO have been solved both in native ... This process operates on the majority of non-halide substrates. The first step is identical followed by: [Fe(IV)=O...Por•+] + ...
Basis for the dual substrate specificity and activation of the oncolytic agent tiazofurin". J. Biol. Chem. 277 (15): 13148-54. ...
"The SPOT technique as a tool for studying protein tyrosine phosphatase substrate specificities". Protein Sci. 11 (10): 2326-34 ...
Goss, R. (2003). "Substrate specificity of the violaxanthin de-epoxidase of the primitive green alga Mantoniella squamata ( ... Lipid composition and substrate specificity". Arch. Biochem. Biophys. 190 (2): 514-522. doi:10.1016/0003-9861(78)90305-3. PMID ...
... the enzyme possesses a high degree of substrate specificity, with the indole moiety of tryptamine required for substrate ... McCoy E, Galan MC, O'Connor SE (2006). "Substrate specificity of strictosidine synthase". Bioorg. Med. Chem. Lett. 16 (9): 2475 ... have suggested that strictosidine synthase can be easily manipulated to have a broader range of substrate specificity. For ... Upon substrate binding, secologanin's position is located at the pocket's entrance, where the positively charged residues His ...
... has broad substrate specificity. EPHX1 detoxifies low molecular weight chemicals, e.g., butadiene, benzene, styrene, etc ... Oesch F (1974). "Purification and specificity of a human microsomal epoxide hydratase". Biochem. J. 139 (1): 77-88. doi:10.1042 ... Androstene oxide and epoxyestratrienol have been shown as endogenous EPHX1 substrates. EPHX1 also metabolizes endocannabinoid 2 ... Structure-activity relationships for substrates and inhibitors". Biochemistry. 10 (26): 4858-66. doi:10.1021/bi00802a005. PMID ...
Substrate specificity and mechanistic implications". J. Biol. Chem. 270 (40): 23540-5. doi:10.1074/jbc.270.40.23540. PMID ... the two substrates of this enzyme are L-tryptophan and O2, whereas its two products are alpha,beta-didehydrotryptophan and H2O2 ...
Domain structure and substrate specificity". The Journal of Biological Chemistry. 275 (15): 10983-8. doi:10.1074/jbc.275.15. ... or they may alter the substrate specificity of nearby sites by steric or electrostatic effects. The protein contains 9 or 14 ... Whelan SA, Dias WB, Thiruneelakantapillai L, Lane MD, Hart GW (Feb 2010). "Regulation of insulin receptor substrate 1 (IRS-1)/ ... but the structure of a binary complex with UDP and a ternary complex with UDP and a peptide substrate has been researched. The ...
Implication for a substrate specificity". The Journal of Biological Chemistry. 276 (12): 9158-65. doi:10.1074/jbc.M009250200. ...
When a substrate is thus "poly-ubiquitinated", it is targeted for destruction by the proteasome. Particular enzymes in the ... ubiquitin/proteasome pathway allow ubiquitination to be directed to some proteins but not others - specificity is gained by ... are targeted for destruction by the ligation of at least four copies of a small peptide called ubiquitin onto a substrate ... coupling targeted proteins to an "E3 ubiquitin ligase". Each E3 ubiquitin ligase binds to a particular set of substrates, ...
Koudelakova T, Chovancova E, Brezovsky J, Monincova M, Fortova A, Jarkovsky J, Damborsky J (2011). "Substrate Specificity of ... To begin, aspartate 124 is perfectly aligned with the substrate. It will drive off the halogen and form an ester functionality ... the two substrates of this enzyme are 1-haloalkane and H2O, whereas its two products are primary alcohol and halide. This ... a dehalogenase of wide substrate range from an Arthrobacter sp". J. Bacteriol. 169 (11): 5016-21. PMC 213902 . PMID 3667524. ...
Molecular determinants of metalloproteinase substrate specificity. Matrix metalloproteinase substrate binding domains, modules ... substrate specificity, and tissue inhibitor of metalloproteinase interaction. J. Biol. Chem. 272, 7608-7616.PubMedCrossRef ... cDNA cloning and deprivation of protein substrate specificity by autolytic degradation. Biochemistry 36, 7225-7238.PubMed ... identification of regions responsible for substrate specificity and general proteinase activity. Proc. Natl. Acad. Sci. USA. 90 ...
... they showed enhanced substrate specificity relative to the native enzyme. This increased specificity was achieved by the ... Redesigning trypsin: alteration of substrate specificity.. Craik CS, Largman C, Fletcher T, Roczniak S, Barr PJ, Fletterick R, ... Computer graphic analysis suggested that these substitutions would differentially affect arginine and lysine substrate binding ... at position 226 exhibited an altered conformation that may be converted to a trypsin-like structure upon binding of a substrate ...
... Andrew M. Slupe, Ronald A. Merrill, and Stefan Strack ... Andrew M. Slupe, Ronald A. Merrill, and Stefan Strack, "Determinants for Substrate Specificity of Protein Phosphatase 2A," ...
Kinetic analysis reveals that the major source of the substrate specificity lies in changes in K(m) for the various substrates ... Substrate specificity of RdgB protein, a deoxyribonucleoside triphosphate pyrophosphohydrolase.. Burgis NE1, Cunningham RP. ... possess deoxyribonucleoside triphosphate pyrophosphohydrolase activity and that all four RdgB homologs have high specificity ...
Anatomical context of Substrate Specificity. *The strict substrate specificity of this reaction suggests that L-arginine is the ... Gene context of Substrate Specificity. *The substrate specificity of the truncated RAD2 protein implicates branched DNA ... Disease relevance of Substrate Specificity. *Altered substrate specificity of herpes simplex virus thymidine kinase confers ... Substrate specificity and affinity of a protein modulated by bound water molecules. Quiocho, F.A., Wilson, D.K., Vyas, N.K. ...
188af) Exploring and Enhancing the Activity and Substrate Specificity of Amine Dehydrogenases. ... have increased activity across the board without affecting substrate. specificity. Finally, we have demonstrated for the first- ... chimeric amine dehydrogenase shows altered substrate specificity compared to. its parent enzymes. Chem Commun. (Camb) 2014, 50 ... ammonia, ketone, and NADH as substrates and release an (R)-amine, NAD+,. and water as products as seen in the reaction scheme ...
A "Silent" Polymorphism in the MDR1 Gene Changes Substrate Specificity. By Chava Kimchi-Sarfaty, Jung Mi Oh, In-Wha Kim, Zuben ... A "Silent" Polymorphism in the MDR1 Gene Changes Substrate Specificity. By Chava Kimchi-Sarfaty, Jung Mi Oh, In-Wha Kim, Zuben ... A Silent Polymorphism in the MDR1 Gene Changes Substrate Specificity Message Subject. (Your Name) has forwarded a page to you ... mammalian membrane transport protein alters the substrate specificity. We hypothesize that when frequent codons are changed to ...
Peptide substrate specificity also depends on interactions of the peptide with the FPP co-substrate 28-30. As these findings ... These results improve the definition of prenyltransferase substrate specificity, test the efficacy of substrate algorithms, and ... Biochemical studies of prenyltransferase substrate specificity indicate that recognition of peptide substrates is more complex ... the MTO substrates, the STO substrates, and the non-substrate peptides. Wedges are labeled with the percentage representation ...
... Laadan, B. ... resulting in Adh1p-variants with different substrate specificities. ... Kinetic characterization with both aldehydes and the in vivo primary substrate acetaldehyde also enabled to correlate the ... of both native and mutated ADH1 genes was performed in order to identify the key amino acids involved in this substrate shift, ...
Japans largest platform for academic e-journals: J-STAGE is a full text database for reviewed academic papers published by Japanese societies
... Valsecchi, Isabel Uppsala University, Disciplinary Domain of Medicine and Pharmacy, ... All substrates were dephosphorylated by AtZDP, assuming that this enzyme could potentially be involved in double-strand DNA ...
Molecular model of the specificity pocket of the hepatitis C virus protease: implications for substrate recognition.. E Pizzi, ... Molecular model of the specificity pocket of the hepatitis C virus protease: implications for substrate recognition. ... Molecular model of the specificity pocket of the hepatitis C virus protease: implications for substrate recognition. ... Molecular model of the specificity pocket of the hepatitis C virus protease: implications for substrate recognition. ...
analogues-exploring the substrate specificity of PigC Suresh R. Chawrai,a Neil R. Williamson,b George P. C. Salmond*b and ... analogues-exploring the substrate specificity of PigC S. R. Chawrai, N. R. Williamson, G. P. C. Salmond and F. J. Leeper, Chem ...
The invention also provides peptide substrates for protein kinase A, cell cycle control kinases, src family kinases, the EGF ... peptides within the library which are substrates for the kinase are converted to phosphopeptides and the phosphopeptides are ... When the substrate is a substrate for a protein-serine/threonine kinase, Xaa5 is Ser or Thr, whereas whenthe substrate is a ... This invention pertains to the substrate specificity of protein kinases and to peptides which are substrates for protein ...
In this work we employed a novel version of substrate phage display (21) to analyze the extended substrate specificity of OmpT ... We have analyzed the substrate specificity of OmpT by two complementary substrate filamentous phage display methods: (i) in ... Substrate Specificity of the Escherichia coli Outer Membrane Protease OmpT. John D. McCarter, Daren Stephens, Kevin Shoemaker, ... Substrate Specificity of the Escherichia coli Outer Membrane Protease OmpT. John D. McCarter, Daren Stephens, Kevin Shoemaker, ...
... Byrne R.T., Jenkins H.T., Peters D.T. ... The exquisite substrate specificity of Dus enzymes is therefore controlled by a relatively simple mechanism involving major ... Such reprogramming of the enzymatic specificity appears to be a unique evolutionary solution for altering tRNA recognition by ... that Dus subfamilies that selectively modify U16 or U20 in tRNA adopt identical folds but bind their respective tRNA substrates ...
Mutations affecting substrate specificity of the Bacillus subtilis multidrug transporter Bmr.. K A Klyachko, S Schuldiner, A A ... Mutations affecting substrate specificity of the Bacillus subtilis multidrug transporter Bmr.. K A Klyachko, S Schuldiner, A A ... Mutations affecting substrate specificity of the Bacillus subtilis multidrug transporter Bmr.. K A Klyachko, S Schuldiner, A A ... Mutations affecting substrate specificity of the Bacillus subtilis multidrug transporter Bmr. Message Subject (Your Name) has ...
Crystal structure of the FTO protein reveals basis for its substrate specificity.. [Zhifu Han, Tianhui Niu, Junbiao Chang, ... Taken together, these results provide a structural basis for understanding FTO substrate-specificity, and serve as a foundation ...
A characteristic feature of enzyme activity in relation to the kind of substrate on which the enzyme or catalytic molecule ... Substrate Specificity. Known as: Specificities, Substrate, Specificity, Substrate, Substrate Specificities A characteristic ... Studies of the substrate specificity, enzyme-substrate interactions, and the function of lipid. ... Substrate specificity and redox potential of AhpC, a bacterial peroxiredoxin.. *Derek Parsonage, P. Andrew Karplus, Leslie B. ...
... Ilari A., ... Accordingly, a 15 fold increase in k(cat) and 100 fold decrease in K(m) for sarcosine as substrate has been achieved in MTOX ... The analysis of the substrate binding site highlights the structural determinants that favour the accommodation of the bulky N- ... located at the entrance of the active site appears to play a key role for the recognition of the amino acid substrate side ...
Substrate Specificity of Acyl-Lipid Δ9-Desaturase from Cyanobacterium sp. IPPAS B-1200, a Cyanobacterium with Unique Fatty Acid ... We cloned this gene and characterized its specificity to the length of the substrate using heterologous expression in Escheri ... Substrate specificity of acyl-lipid Δ9-desaturase from Prochlorothrix hollandica cyanobacterium producing myristoleic acid, ...
... thereby linking the substrate specificity to the known host tropism. The novel IgdE family proteases of S. agalactiae, S. ... Porcine IgG subtype specificities of the IgdE family proteases of S. porcinus and S. pseudoporcinus remain to be determined. ... The IgG subtype specificity of bacterial proteases indicates the special importance of these IgG subtypes in counteracting ... pseudoporcinus and S. equi showed IgG subtype specificity, i.e. IgdE from S. agalactiae and S. pseudoporcinus cleaved human ...
... and modulate substrate specificity. Two amino acid differences at the domain interface alter the substrate binding specificity ... To avoid cleavage of unintended substrates, these enzymes have very selective substrate specificities. Our biochemical and ... Title: Insights into substrate specificity of NlpC/P60 cell wall hydrolases containing bacterial SH3 domains ... Accepted Manuscript: Insights into substrate specificity of NlpC/P60 cell wall hydrolases containing bacterial SH3 domains ...
Highlights • The origin of specificity of the LeuRS editing domain was until now unresolved. • Ground-state binding does not... ... Kinetic Origin of Substrate Specificity in Post-Transfer Editing by Leucyl-tRNA Synthetase. ... Kinetic Origin of Substrate Specificity in Post-Transfer Editing by Leucyl-tRNA Synthetase ... Kinetic Origin of Substrate Specificity in Post-Transfer Editing by Leucyl-tRNA Synthetase ...
... lipases are poorly characterized with regard to their substrate specificities and physiological functions. Here ... ... Presently, it seems more feasible to explore substrate specificities for groups of enzymes where at least one substrate is ... Defining Substrate Specificities for Lipase and Phospholipase Candidates. Diana X. Sahonero-Canavesi1, Maritza Zavaleta-Pastor1 ... The definition of substrate specificities for new enzymes, often provides hypotheses for a potential physiological role of ...
  • 1991) "Peptide-Binding Specificity of the Molecular Chaperone BiP" Nature 353:726-730. (patentgenius.com)
  • Finally, using molecular dynamics, we generate a model of the Michaelis complex of the substrate bound in the active site of GlpG. (embopress.org)
  • Structure‐based modeling and molecular dynamics simulations allow generating a model of the Michaelis complex with the substrate. (embopress.org)
  • A substrate-inhibitory analysis with the use of deprenyl and chloroginyl provides an indirect evidence for the existence of a sole MAO molecular form in the Kamchatka crab hepatopancreas. (springer.com)
  • The substrate-specific impact of residue 196 was studied by molecular dynamics simulations using a cationic dummy atom approach for the zinc ions. (caltech.edu)
  • To model stability, six molecular dynamics simulations at 100 K were carried out for all enzymesubstrate complexes. (caltech.edu)
  • The results evaluated are in good agreement with available experimental data, thus providing detailed molecular models which can explain the structural and specificities differences between both zinc peptidases. (biomedcentral.com)
  • The model allowed us to predict that the substrate of this protease should have a cysteine residue in position P1. (pnas.org)
  • Super-reactive' substrate sequences were engineered, with more than a 2-fold increase in activity, by combining the best residue choices at P5 to P3 positions. (hkmj.org)
  • This change in specificity occurs even though residue 262 is remote from the active site. (caltech.edu)
  • We investigated the substrate specificities of five other point mutants resulting from single-nucleotide substitutions at positions near residue 262: G262A, G262V, S121G, F218Y, and F218I. (caltech.edu)
  • Moreover, besides the effect of substrate entrance on its own, we strongly suggest that a highly conserved arginine residue (in the RDP motif) rather than the previously proposed Tyr motif (not conserved) provides the proton to increase the pKa of the acid-base catalyst. (kuleuven.be)
  • The results suggested that residue Leu199 of DMWf18-543 shortens and blocks the substrate-binding pocket. (biomedcentral.com)
  • Nevertheless, the enzyme's high thermostability and broad acceptor specificity makes it a valuable candidate for industrial disaccharide synthesis. (ugent.be)
  • As the size of the ('3)H-oligosaccharide acceptor increases, the maximum velocity of the enzyme increases for chondroitin-6-sulfate and chondroitin substrates. (illinois.edu)
  • Probing the substrate specificity of Trypanosom. (lancs.ac.uk)
  • Vascular peroxidase 1 catalyzes the formation of hypohalous acids: characterization of its substrate specificity and enzymatic properties. (biomedsearch.com)
  • Enzymatic properties and substrate sp. (ugent.be)
  • Van der Borght J, Chen C, Hoflack L, Van Renterghem L, Desmet T, Soetaert W. Enzymatic properties and substrate specificity of the trehalose phosphorylase from Caldanaerobacter subterraneus. (ugent.be)
  • Many important experiments in proteomics including protein digestion, enzyme substrate screening, enzymatic labeling, etc., involve the enzymatic reactions in a complex system where numerous substrates coexists with an enzyme. (mcponline.org)
  • In such experiments, an enzyme was incubated with numerous competing substrates for enzymatic reaction which would generate numerous different products. (mcponline.org)
  • Herein, we demonstrate that a bacterial PAL from Anabaena variabilis (AvPAL) displays significantly higher activity towards a series of non-natural substrates than previously described eukaryotic PALs. (manchester.ac.uk)
  • The substrate specificity of MT is significantly affected by the protein context within which it is present. (biomedcentral.com)
  • We have built a model of the specificity pocket of the protease of hepatitis C virus on the basis of the known structures of trypsin-like serine proteases and of the conservation pattern of the protease sequences among various hepatitis C strains. (pnas.org)
  • The mechanisms of intramembrane proteases are incompletely understood due to the lack of structural data on substrate complexes. (embopress.org)
  • Despite valuable insights from inhibitor‐bound rhomboid proteases, full understanding of intramembrane proteolysis requires structural views of protease‐substrate complexes. (embopress.org)
  • Here we present a detailed study of substrate preferences of all three proteases. (uni-bielefeld.de)
  • 2012). Substrate specificity of Staphylococcus aureus cysteine proteases - Staphopains A, B and C. Biochimie , 94 (2), 318-327. (uni-bielefeld.de)
  • Substrate specificity of Staphylococcus aureus cysteine proteases - Staphopains A, B and C". Biochimie 94.2 (2012): 318-327. (uni-bielefeld.de)
  • This improved understanding of EaDAcT specificity confirms that the enzyme preferentially utilizes acetyl-CoA to acetylate sn-1,2-DAGs and will be helpful in engineering the production of acetyl-TAG with improved functionality in transgenic plants. (k-state.edu)
  • Herein, we demonstrate that the microenvironment from which Ras signals emanate determines which substrates will be preferentially phosphorylated by the activated ERK1/2. (hud.ac.uk)
  • Substrate preferences at P5 to P3 positions were important in enhancing the main protease activity. (hkmj.org)
  • We show that peptidyl‐CMKs derived from the natural rhomboid substrate TatA from bacterium Providencia stuartii bind GlpG in a substrate‐like manner, and their co‐crystal structures with GlpG reveal the S1 to S4 subsites of the protease. (embopress.org)
  • Here we show that RdgB protein and RdgB homologs from Saccharomyces cerevisiae, mouse, and human all possess deoxyribonucleoside triphosphate pyrophosphohydrolase activity and that all four RdgB homologs have high specificity for dHAPTP and deoxyinosine triphosphate compared with the four canonical dNTPs and several other noncanonical (d)NTPs. (nih.gov)
  • In most cases the results show flexibility in the P4 position, very high specificity for arginine in the P3 position and glycine in the P2 position, in accord with the sequence of the natural substrate, ubiquitin. (portlandpress.com)
  • This hypothesis was confirmed by the finding that the DMWf18-558-A199L mutant showed a similar substrate specificity profile to that of DMWf18-543. (biomedcentral.com)
  • The results suggest that a modification of PKC alpha in situ limits its substrate specificity, and indicate an additional level of control of the kinase that may be a site for modulation of PKC-mediated signal transduction. (garvan.org.au)
  • The analysis demonstrated that the most efficiently hydrolyzed sites, using Schechter and Berger nomenclature, comprise a P2-Gly down arrow Ala(Ser) sequence motif, where P2 distinguishes the specificity of staphopain A (Leu) from that of both staphopains B and C (Phe/Tyr). (uni-bielefeld.de)
  • Schnepel J, Unger J, Tschesche H. Recombinant cryptic human fibronectinase cleaves actin and myosin: Substrate specificity and possible role in muscular dystrophy. (uni-bielefeld.de)
  • The results described herein are a first step toward the systematic evaluation of a panel of dog P450s and the development of dog P450 isoenzyme-selective marker substrates, as well as providing useful information on prediction and extrapolation of the results from in vitro to in vivo and from dog to human. (aspetjournals.org)
  • The 6-Foot Mini Toober Activity shows students, through modeling, how even a slight change in the shape of either the substrate or the enzyme may alter the efficient and selective ability of the enzyme to catalyze the reaction. (3dmoleculardesigns.com)
  • These results provide a basis for the engineering of mammalian STs to accommodate non-natural substrate analogs that should prove valuable as chemical biological probes of sialyltransferase function. (ubc.ca)
  • X-converting enzyme (XCE) involved in nervous control of respiration, is a member of the M13 family of zinc peptidases, for which no natural substrate has been identified yet. (biomedcentral.com)