Biotinylation: Incorporation of biotinyl groups into molecules.Biotin: A water-soluble, enzyme co-factor present in minute amounts in every living cell. It occurs mainly bound to proteins or polypeptides and is abundant in liver, kidney, pancreas, yeast, and milk.Carbon-Nitrogen Ligases: Enzymes that catalyze the joining of two molecules by the formation of a carbon-nitrogen bond. EC 6.3.Biotinidase: An enzyme which catalyzes the release of BIOTIN from biocytin. In human, defects in the enzyme are the cause of the organic acidemia MULTIPLE CARBOXYLASE DEFICIENCY or BIOTINIDASE DEFICIENCY.Streptavidin: A 60-kDa extracellular protein of Streptomyces avidinii with four high-affinity biotin binding sites. Unlike AVIDIN, streptavidin has a near neutral isoelectric point and is free of carbohydrate side chains.Avidin: A specific protein in egg albumin that interacts with BIOTIN to render it unavailable to mammals, thereby producing biotin deficiency.Fatty Acid Synthase, Type II: The form of fatty acid synthase complex found in BACTERIA; FUNGI; and PLANTS. Catalytic steps are like the animal form but the protein structure is different with dissociated enzymes encoded by separate genes. It is a target of some ANTI-INFECTIVE AGENTS which result in disruption of the CELL MEMBRANE and CELL WALL.Cell Membrane: The lipid- and protein-containing, selectively permeable membrane that surrounds the cytoplasm in prokaryotic and eukaryotic cells.Methylmalonyl-CoA Decarboxylase: A carboxy-lyase that catalyzes the decarboxylation of (S)-2-Methyl-3-oxopropanoyl-CoA to propanoyl-CoA. In microorganisms the reaction can be coupled to the vectorial transport of SODIUM ions across the cytoplasmic membrane.Holocarboxylase Synthetase Deficiency: The neonatal form of MULTIPLE CARBOXYLASE DEFICIENCY that is caused by a defect or deficiency in holocarboxylase synthetase. HLCS is the enzyme that covalently links biotin to the biotin dependent carboxylases (propionyl-CoA-carboxylase, pyruvate carboxylase, and beta-methylcrotonyl-CoA carboxylase).Protein Transport: The process of moving proteins from one cellular compartment (including extracellular) to another by various sorting and transport mechanisms such as gated transport, protein translocation, and vesicular transport.Cell Line: Established cell cultures that have the potential to propagate indefinitely.Carboxyl and Carbamoyl Transferases: A group of enzymes that catalyze the transfer of carboxyl- or carbamoyl- groups. EC 2.1.3.Carbon-Carbon Ligases: Enzymes that catalyze the joining of two molecules by the formation of a carbon-carbon bond. These are the carboxylating enzymes and are mostly biotinyl-proteins. EC 6.4.Acetyl-CoA Carboxylase: A carboxylating enzyme that catalyzes the conversion of ATP, acetyl-CoA, and HCO3- to ADP, orthophosphate, and malonyl-CoA. It is a biotinyl-protein that also catalyzes transcarboxylation. The plant enzyme also carboxylates propanoyl-CoA and butanoyl-CoA (From Enzyme Nomenclature, 1992) EC 6.4.1.2.Epigenetic Repression: The turning off of GENETIC TRANSCRIPTION in certain regions of CHROMATIN without changes in the DNA sequence. Typically epigenetic repression is a way that developmental changes are programmed at the cellular level.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.Multiple Carboxylase Deficiency: A deficiency in the activities of biotin-dependent enzymes (propionyl-CoA carboxylase, methylcrotonyl-CoA carboxylase, and PYRUVATE CARBOXYLASE) due to one of two defects in BIOTIN metabolism. The neonatal form is due to HOLOCARBOXYLASE SYNTHETASE DEFICIENCY. The late-onset form is due to BIOTINIDASE DEFICIENCY.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.Carbonic Anhydrase I: A cytosolic carbonic anhydrase isoenzyme primarily expressed in ERYTHROCYTES, vascular endothelial cells, and the gastrointestinal mucosa. EC 4.2.1.-Endocytosis: Cellular uptake of extracellular materials within membrane-limited vacuoles or microvesicles. ENDOSOMES play a central role in endocytosis.Lysine: An essential amino acid. It is often added to animal feed.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.Histones: Small chromosomal proteins (approx 12-20 kD) possessing an open, unfolded structure and attached to the DNA in cell nuclei by ionic linkages. Classification into the various types (designated histone I, histone II, etc.) is based on the relative amounts of arginine and lysine in each.Succinimides: A subclass of IMIDES with the general structure of pyrrolidinedione. They are prepared by the distillation of ammonium succinate. They are sweet-tasting compounds that are used as chemical intermediates and plant growth stimulants.Pyruvate Carboxylase: A biotin-dependent enzyme belonging to the ligase family that catalyzes the addition of CARBON DIOXIDE to pyruvate. It is occurs in both plants and animals. Deficiency of this enzyme causes severe psychomotor retardation and ACIDOSIS, LACTIC in infants. EC 6.4.1.1.Symporters: Membrane transporters that co-transport two or more dissimilar molecules in the same direction across a membrane. Usually the transport of one ion or molecule is against its electrochemical gradient and is "powered" by the movement of another ion or molecule with its electrochemical gradient.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.Escherichia coli Proteins: Proteins obtained from ESCHERICHIA COLI.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.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.HEK293 Cells: A cell line generated from human embryonic kidney cells that were transformed with human adenovirus type 5.Repressor Proteins: Proteins which maintain the transcriptional quiescence of specific GENES or OPERONS. Classical repressor proteins are DNA-binding proteins that are normally bound to the OPERATOR REGION of an operon, or the ENHANCER SEQUENCES of a gene until a signal occurs that causes their release.Cell Polarity: Orientation of intracellular structures especially with respect to the apical and basolateral domains of the plasma membrane. Polarized cells must direct proteins from the Golgi apparatus to the appropriate domain since tight junctions prevent proteins from diffusing between the two domains.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.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.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.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.Microscopy, Confocal: A light microscopic technique in which only a small spot is illuminated and observed at a time. An image is constructed through point-by-point scanning of the field in this manner. Light sources may be conventional or laser, and fluorescence or transmitted observations are possible.Cricetinae: A subfamily in the family MURIDAE, comprising the hamsters. Four of the more common genera are Cricetus, CRICETULUS; MESOCRICETUS; and PHODOPUS.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.Dogs: The domestic dog, Canis familiaris, comprising about 400 breeds, of the carnivore family CANIDAE. They are worldwide in distribution and live in association with people. (Walker's Mammals of the World, 5th ed, p1065)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.Immunoprecipitation: The aggregation of soluble ANTIGENS with ANTIBODIES, alone or with antibody binding factors such as ANTI-ANTIBODIES or STAPHYLOCOCCAL PROTEIN A, into complexes large enough to fall out of solution.Staining and Labeling: The marking of biological material with a dye or other reagent for the purpose of identifying and quantitating components of tissues, cells or their extracts.Pyrococcus horikoshii: Anaerobic hyperthermophilic species of ARCHAEA, isolated from hydrothermal fluid samples. It is obligately heterotrophic with coccoid cells that require TRYPTOPHAN for growth.Mass Spectrometry: An analytical method used in determining the identity of a chemical based on its mass using mass analyzers/mass spectrometers.Glutamate Plasma Membrane Transport Proteins: A family of plasma membrane neurotransmitter transporter proteins that couple the uptake of GLUTAMATE with the import of SODIUM ions and PROTONS and the export of POTASSIUM ions. In the CENTRAL NERVOUS SYSTEM they regulate neurotransmission through synaptic reuptake of the excitatory neurotransmitter glutamate. Outside the central nervous system they function as signal mediators and regulators of glutamate metabolism.Kidney Tubules, Collecting: Straight tubes commencing in the radiate part of the kidney cortex where they receive the curved ends of the distal convoluted tubules. In the medulla the collecting tubules of each pyramid converge to join a central tube (duct of Bellini) which opens on the summit of the papilla.CHO Cells: CELL LINE derived from the ovary of the Chinese hamster, Cricetulus griseus (CRICETULUS). The species is a favorite for cytogenetic studies because of its small chromosome number. The cell line has provided model systems for the study of genetic alterations in cultured mammalian cells.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.Carrier Proteins: Transport proteins that carry specific substances in the blood or across cell membranes.Solute Carrier Family 12, Member 1: Na-K-Cl transporter in the ASCENDING LIMB OF LOOP OF HENLE. It mediates active reabsorption of sodium chloride and is inhibited by LOOP DIURETICS such as FUROSEMIDE; and BUMETANIDE. Mutations in the gene encoding SLC12A1 are associated with a BARTTER SYNDROME.Epithelial Sodium Channels: Sodium channels found on salt-reabsorbing EPITHELIAL CELLS that line the distal NEPHRON; the distal COLON; SALIVARY DUCTS; SWEAT GLANDS; and the LUNG. They are AMILORIDE-sensitive and play a critical role in the control of sodium balance, BLOOD VOLUME, and BLOOD PRESSURE.Propionibacterium: A genus of gram-positive, rod-shaped bacteria whose cells occur singly, in pairs or short chains, in V or Y configurations, or in clumps resembling letters of the Chinese alphabet. Its organisms are found in cheese and dairy products as well as on human skin and can occasionally cause soft tissue infections.Magnetospirillum: A genus of microaerophilic, gram-negative bacteria that forms crystals of the mineral magnetite in special organelles called MAGNETOSOMES.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.Microscopy, Fluorescence: Microscopy of specimens stained with fluorescent dye (usually fluorescein isothiocyanate) or of naturally fluorescent materials, which emit light when exposed to ultraviolet or blue light. Immunofluorescence microscopy utilizes antibodies that are labeled with fluorescent dye.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.Binding Sites: The parts of a macromolecule that directly participate in its specific combination with another molecule.Epithelial Cells: Cells that line the inner and outer surfaces of the body by forming cellular layers (EPITHELIUM) or masses. Epithelial cells lining the SKIN; the MOUTH; the NOSE; and the ANAL CANAL derive from ectoderm; those lining the RESPIRATORY SYSTEM and the DIGESTIVE SYSTEM derive from endoderm; others (CARDIOVASCULAR SYSTEM and LYMPHATIC SYSTEM) derive from mesoderm. Epithelial cells can be classified mainly by cell shape and function into squamous, glandular and transitional epithelial cells.Carboxy-Lyases: Enzymes that catalyze the addition of a carboxyl group to a compound (carboxylases) or the removal of a carboxyl group from a compound (decarboxylases). EC 4.1.1.Jurkat Cells: A CELL LINE derived from human T-CELL LEUKEMIA and used to determine the mechanism of differential susceptibility to anti-cancer drugs and radiation.Mutagenesis, Site-Directed: Genetically engineered MUTAGENESIS at a specific site in the DNA molecule that introduces a base substitution, or an insertion or deletion.Cystic Fibrosis Transmembrane Conductance Regulator: A chloride channel that regulates secretion in many exocrine tissues. Abnormalities in the CFTR gene have been shown to cause cystic fibrosis. (Hum Genet 1994;93(4):364-8)Recombinant Proteins: Proteins prepared by recombinant DNA technology.Opossums: New World marsupials of the family Didelphidae. Opossums are omnivorous, largely nocturnal and arboreal MAMMALS, grow to about three feet in length, including the scaly prehensile tail, and have an abdominal pouch in which the young are carried at birth.Dopamine Plasma Membrane Transport Proteins: Sodium chloride-dependent neurotransmitter symporters located primarily on the PLASMA MEMBRANE of dopaminergic neurons. They remove DOPAMINE from the EXTRACELLULAR SPACE by high affinity reuptake into PRESYNAPTIC TERMINALS and are the target of DOPAMINE UPTAKE INHIBITORS.Kidney: Body organ that filters blood for the secretion of URINE and that regulates ion concentrations.Caco-2 Cells: Human colonic ADENOCARCINOMA cells that are able to express differentiation features characteristic of mature intestinal cells, such as ENTEROCYTES. These cells are valuable in vitro tools for studies related to intestinal cell function and differentiation.Brefeldin A: A fungal metabolite which is a macrocyclic lactone exhibiting a wide range of antibiotic activity.Immunoblotting: Immunologic method used for detecting or quantifying immunoreactive substances. The substance is identified by first immobilizing it by blotting onto a membrane and then tagging it with labeled antibodies.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)Cysteine: A thiol-containing non-essential amino acid that is oxidized to form CYSTINE.Bacterial Proteins: Proteins found in any species of bacterium.

A single membrane-embedded negative charge is critical for recognizing positively charged drugs by the Escherichia coli multidrug resistance protein MdfA. (1/1806)

The nature of the broad substrate specificity phenomenon, as manifested by multidrug resistance proteins, is not yet understood. In the Escherichia coli multidrug transporter, MdfA, the hydrophobicity profile and PhoA fusion analysis have so far identified only one membrane-embedded charged amino acid residue (E26). In order to determine whether this negatively charged residue may play a role in multidrug recognition, we evaluated the expression and function of MdfA constructs mutated at this position. Replacing E26 with the positively charged residue lysine abolished the multidrug resistance activity against positively charged drugs, but retained chloramphenicol efflux and resistance. In contrast, when the negative charge was preserved in a mutant with aspartate instead of E26, chloramphenicol recognition and transport were drastically inhibited; however, the mutant exhibited almost wild-type multidrug resistance activity against lipophilic cations. These results suggest that although the negative charge at position 26 is not essential for active transport, it dictates the multidrug resistance character of MdfA. We show that such a negative charge is also found in other drug resistance transporters, and its possible significance regarding multidrug resistance is discussed.  (+info)

Comparison of the backbone dynamics of the apo- and holo-carboxy-terminal domain of the biotin carboxyl carrier subunit of Escherichia coli acetyl-CoA carboxylase. (2/1806)

The biotin carboxyl carrier protein (BCCP) is a subunit of acetyl-CoA carboxylase, a biotin-dependent enzyme that catalyzes the first committed step of fatty acid biosynthesis. In its functional cycle, this protein engages in heterologous protein-protein interactions with three distinct partners, depending on its state of post-translational modification. Apo-BCCP interacts specifically with the biotin holoenzyme synthetase, BirA, which results in the post-translational attachment of biotin to a single lysine residue on BCCP. Holo-BCCP then interacts with the biotin carboxylase subunit of acetyl-CoA carboxylase, which leads to the addition of the carboxylate group of bicarbonate to biotin. Finally, the carboxy-biotinylated form of BCCP interacts with transcarboxylase in the transfer of the carboxylate to acetyl-CoA to form malonyl-CoA. The determinants of protein-protein interaction specificity in this system are unknown. The NMR solution structure of the unbiotinylated form of an 87 residue C-terminal domain fragment (residue 70-156) of BCCP (holoBCCP87) and the crystal structure of the biotinylated form of a C-terminal fragment (residue 77-156) of BCCP from Escherichia coli acetyl-CoA carboxylase have previously been determined. Comparative analysis of these structures provided evidence for small, localized conformational changes in the biotin-binding region upon biotinylation of the protein. These structural changes may be important for regulating specific protein-protein interactions. Since the dynamic properties of proteins are correlated with local structural environments, we have determined the relaxation parameters of the backbone 15N nuclear spins of holoBCCP87, and compared these with the data obtained for the apo protein. The results indicate that upon biotinylation, the inherent mobility of the biotin-binding region and the protruding thumb, with which the biotin group interacts in the holo protein, are significantly reduced.  (+info)

Streptavidin facilitates internalization and pulmonary targeting of an anti-endothelial cell antibody (platelet-endothelial cell adhesion molecule 1): a strategy for vascular immunotargeting of drugs. (3/1806)

Conjugation of drugs with antibodies to surface endothelial antigens is a potential strategy for drug delivery to endothelium. We studied antibodies to platelet-endothelial adhesion molecule 1 (PECAM-1, a stably expressed endothelial antigen) as carriers for vascular immunotargeting. Although 125I-labeled anti-PECAM bound to endothelial cells in culture, the antibody was poorly internalized by the cells and accumulated poorly after intravenous administration in mice and rats. However, conjugation of biotinylated anti-PECAM (b-anti-PECAM) with streptavidin (SA) markedly stimulated uptake and internalization of anti-PECAM by endothelial cells and by cells expressing PECAM. In addition, conjugation with streptavidin markedly stimulated uptake of 125I-labeled b-anti-PECAM in perfused rat lungs and in the lungs of intact animals after either intravenous or intraarterial injection. The antioxidant enzyme catalase conjugated with b-anti-PECAM/SA bound to endothelial cells in culture, entered the cells, escaped intracellular degradation, and protected the cells against H2O2-induced injury. Anti-PECAM/SA/125I-catalase accumulated in the lungs after intravenous injection or in the perfused rat lungs and protected these lungs against H2O2-induced injury. Thus, modification of a poor carrier antibody with biotin and SA provides an approach for facilitation of antibody-mediated drug targeting. Anti-PECAM/SA is a promising candidate for vascular immunotargeting of bioactive drugs.  (+info)

Molecular biology of biotin attachment to proteins. (4/1806)

Enzymatic attachment of biotin to proteins requires the interaction of a distinct domain of the acceptor protein (the "biotin domain") with the enzyme, biotin protein ligase, that catalyzes this essential and rare post-translational modification. Both biotin domains and biotin protein ligases are very strongly conserved throughout biology. This review concerns the protein structures and mechanisms involved in the covalent attachment of biotin to proteins.  (+info)

Human biotinidase isn't just for recycling biotin. (5/1806)

For years, the major role of biotin has been as the coenzyme for four carboxylases in humans. Although there has been evidence that biotin might have other functions, none has been firmly established. The discovery that human serum biotinidase has biotinyl-transferase activity, in addition to biotinidase hydrolase activity, presents new possibilities for the role of biotinidase in biotin metabolism. Specific transfer of biotin to histones by biotinidase provides a possible explanation for why biotin is found in the nucleus and the nature of its role in the regulation of protein transcription. Future studies will help to determine the functions of biotinidase in biotin metabolism and in disease states.  (+info)

Treatment of mouse oocytes with PI-PLC releases 70-kDa (pI 5) and 35- to 45-kDa (pI 5.5) protein clusters from the egg surface and inhibits sperm-oolemma binding and fusion. (6/1806)

The effect of phosphatidyinositol-specific phospholipase C (PI-PLC) on mouse sperm-egg interaction was investigated in this study to determine if glycosyl-phosphatidylinositol (GPI)-anchored proteins are involved in mammalian fertilization. When both sperm and zona-intact oocytes were pretreated with a highly purified preparation of PI-PLC and coincubated, there was no significant effect on sperm-zona pellucida binding; however, fertilization was reduced from 59.6% (control group) to 2.8% (treatment group). A similar reduction in fertilization rates was found when zona-intact oocytes were treated with PI-PLC and washed prior to incubation with untreated sperm. The effect of PI-PLC on sperm binding and fusion with zona-free oocytes was then investigated. Treatment of sperm with PI-PLC had no significant effect on sperm-egg binding or fusion. However, treatment of eggs with PI-PLC significantly reduced sperm-egg binding and fusion from 6.2 bound and 2.1 fused sperm per egg in the control group to 2.1 bound and 0.02 fused sperm per egg in the treatment group. This decrease in sperm-egg binding and fusion depended on the dose of PI-PLC employed, with a maximal inhibitory effect on binding and fusion at 5 and 1 U/ml, respectively. PI-PLC-treated oocytes could be artificially activated by calcium ionophore, demonstrating that the oocytes were functionally viable following treatment. Furthermore, treatment of oocytes with PI-PLC did not reduce the immunoreactivity of the non-GPI-anchored egg surface integrin, alpha6beta1. Taken together, these observations support the hypothesis that PI-PLC affects fertilization by specifically releasing GPI-anchored proteins from the oolemma. In order to identify the oolemmal GPI-anchored proteins involved in fertilization, egg surface proteins were labeled with sulfo-NHS biotin, treated with PI-PLC, and analyzed by two-dimensional gel electrophoresis followed by avidin blotting. A prominent high-molecular-weight protein cluster (approximately 70 kDa, pI 5) and a lower molecular weight (approximately 35-45 kDa, pI 5.5) protein cluster were released from the oolemmal surface as a result of PI-PLC treatment. It is likely that these GPI-anchored egg surface proteins are required for sperm-egg binding and fusion.  (+info)

Critical relationship between glycosylation of recombinant lutropin receptor ectodomain and its secretion from baculovirus-infected insect cells. (7/1806)

The lutropin receptor ectodomain overexpressed under the control of the powerful polyhedrin promoter in baculovirus-infected Sf9 insect cells, is mainly found in an inactive, intracellularly-aggregated form. It is secreted in an active form under the control of the P10 promoter, a somewhat weaker and earlier promoter, at the price of a lower production. The apparent molecular masses of the two species encoded by the same cDNA are 48 kDa and 60-68 kDa, respectively. The relationship between the extent and type of glycosylation and the extracellular targeting for the recombinant lutropin receptor ectodomains was investigated precisely with endoglycosidases, lectins of various specificities, and a glycosylation inhibitor, and tested with monoclonal and polyclonal antibodies. The results indicate that the strong polyhedrin promoter probably overwhelms the processing capacity of the ER in Sf9 cells, so that only a high-mannose precursor is expressed in large amounts. Only a minute amount of protein is secreted, which has been processed by Sf9 exoglycosidases/glycosyltransferases and bears complex/hybrid oligosaccharides. The weaker P10 promoter allows secretion of a mature and active receptor ectodomain, bearing complex glycosylation. An important O-linked glycosylation is also added post-translationally on this species. In particular, beta-galactose and sialic acid residues were specifically detected in the secreted species, evidence of the induction of the corresponding glycosyltransferases or of their genes. These results suggest that Sf9 cells should eventually be engineered with chaperones and glycosyltransferases in order to improve the production of demanding glycoproteins such as the porcine lutropin ectodomain, so as to open the way to resolution of the three-dimensional structures of these receptors.  (+info)

Characterization of the internalization pathways for the cystic fibrosis transmembrane conductance regulator. (8/1806)

Mutations in the gene encoding the cystic fibrosis (CF) transmembrane conductance regulator (CFTR) chloride channel give rise to the most common lethal genetic disease of Caucasian populations, CF. Although the function of CFTR is primarily related to the regulation of apical membrane chloride permeability, biochemical, immunocytochemical, and functional studies indicate that CFTR is also present in endosomal and trans Golgi compartments. The molecular pathways by which CFTR is internalized into intracellular compartments are not fully understood. To define the pathways for CFTR internalization, we investigated the association of CFTR with two specialized domains of the plasma membrane, clathrin-coated pits and caveolae. Internalization of CFTR was monitored after cell surface biotinylation and quantitation of cell surface CFTR levels after elution of cell lysates from a monomeric avidin column. Cell surface levels of CFTR were determined after disruption of caveolae or clathrin-coated vesicle formation. Biochemical assays revealed that disrupting the formation of clathrin-coated vesicles inhibited the internalization of CFTR from the plasma membrane, resulting in a threefold increase in the steady-state levels of cell surface CFTR. In contrast, the levels of cell surface CFTR after disruption of caveolae were not different from those in control cells. In addition, although our studies show the presence of caveolin at the apical membrane domain of human airway epithelial cells, we were unable to detect CFTR in purified caveolae. These results suggest that CFTR is constitutively internalized from the apical plasma membrane via clathrin-coated pits and that CFTR is excluded from caveolae.  (+info)

Target carboxyl groups for biotin labeling using amine-derivatized biotin molecules. Choose the best carboxyl biotinylation reagents for your application!
... enable isolation, separation, concentration and further downstream processing or analysis of biomolecules.
This unit describes a system for expression of biotinylated proteins in mammalian cells in vivo, and its application to chromatin immunoprecipitation (ChIP)
Since its discovery in the first half of the twentieth century, the high‐affinity, noncovalent interaction between biotin (vitamin H) and the avian protein avidin (and its bacterial homologs) has been exploited for many diverse biotechnology applications
Since its discovery in the first half of the twentieth century, the high‐affinity, noncovalent interaction between biotin (vitamin H) and the avian protein avidin (and its bacterial homologs) has been exploited for many diverse biotechnology applications
Project SummaryDuring embryogenesis or wound healing, neighboring cells maintain contact and migrate collectively, thoughthe roles of cell-cell adhesion during collective cell migration is poorly defined. Due to constant pulling andpushing between migrating neighboring cells, we hypothesize that mechanical forces regulate theinteraction between the cell-cell adhesion complex and the actin cytoskeleton, and therefore, theadhesive strength. To identify force-sensitive protein complexes at cell-cell junctions, our innovativebiochemical analysis combines in situ proximal biotin labeling with a cell stretch device that promotes theformation of force-sensitive complexes. By fusing ?-catenin with a promiscuous biotin ligase, any proximalproteins of ?-catenin will be biotinylated. The force-dependent change in the biotinylation profile is an indicationof altered protein complexes. Our preliminary study demonstrates that ?-catenin and myosin IIA are likelyinteracting in a force-dependent manner. While ...
Surface biotinylation and NeutrAvidin pull-down were performed similar to what has been described previously (Zha et al., 2009a; Jing et al., 2011). For biotinylation of organotypic hippocampal slices, each filter, which contained 6-8 slices, was cut out and put in a six-well plate. CHO cells or slices were then washed three times with ice-cold PBS+/+, followed by 30 min incubation at 4°C in 1.5 ml (for CHO cells in 60 mm dishes) or 1 ml (for slices in each well of the 6-well plate) of PBS+/+ containing 0.5 mg/ml Sulfo-NHS-LC-Biotin. Cells were washed once with cold PBS+/+ and the reaction was quenched by 100 mm glycine in PBS+/+. Of note, it is essential, especially for biotinylation of slices, to keep the solution and plates ice cold during the whole procedure. Cells were lysed in 300 μl of NeutrAvidin lysis buffer (PBS, 1% Triton, 0.5% SDS, 0.5 mg/ml N-ethylmaleimide, with protease inhibiters). Cell lysates were sonicated briefly and centrifuged at full speed with a desktop centrifuge for ...
Biotinylated histone H3 peptide acetylated methylated phosphorylated for use in enzyme assays, histone binding and pulldown experiments
Biotinylated histone H2A peptide acetylated methylated phosphorylated for use in enzyme assays, histone binding and pulldown experiments
Abstract: Abstract Background Glioblastoma multiformae (GBM) is the most aggressive type of malignant brain tumor with complex molecular profile. Overexpression of Na+/H+ Exchanger isoform 9 (NHE9) promotes tumor progression and correlates positively with insensitivity to radiochemotherapy and poor prognosis. However, molecular mechanisms responsible for increase in NHE9 levels beyond a critical threshold have not been identified. Methods Bioinformatics analysis, luciferase reporter assays, real-time PCR and western blotting were conducted to examine the expression profiles and identify microRNAs (miRNA) that target NHE9. Cell proliferation and migration assays were conducted in U87 glioblastoma cells to determine the consequence of miRNA mediated targeting of NHE9. Endosomal pH measurements, immunofluorescence microscopy and surface biotinylation experiments were conducted to characterize the mechanistic basis of regulation. Results We show that microRNA 135a (miR-135a) targets NHE9 to ...
We performed bio-ChIP-seq using streptavidin beads immunoprecipitation of biotinylated 5 cardiac transcription factors (fbio) and P300 antibody ChIP-seq. We used a dox-inducible dual adenovirus system to express biotinylated Core Cardiac TFs in HL1. One adenovirus expressed the dox-activated reverse tet activator protein (rtTA) and the E. coli biotinylating enzyme BirA from the cardiac specific rat troponin T promoter. The second virus expressed a Core Cardiac TF fused to a C-terminal flag-bio epitope tag, where bio is the 15 amino acid substrate for BirA. This in vivo biotinylation approach permits pulldown of different factors to be performed under identical, stringent conditions, and circumvents limitations posed by available antibodies. We titrated adenovirus and dox doses to express GATA4flbio and MEF2Aflbio at near endogenous levels. The same conditions were then used to express SRFflbio, TBX5flbio, and NKX2-5flbio. Reference: 12. de Boer E, Rodriguez P, Bonte E, Krijgsveld J, Katsantoni E ...
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TY - JOUR. T1 - CCL2 promotes P2X4 receptor trafficking to the cell surface of microglia. AU - Toyomitsu, Emika. AU - Tsuda, Makoto. AU - Yamashita, Tomohiro. AU - Tozaki-Saitoh, Hidetoshi. AU - Tanaka, Yoshitaka. AU - Inoue, Kazuhide. PY - 2012/6/1. Y1 - 2012/6/1. N2 - P2X4 receptors (P2X4Rs), a subtype of the purinergic P2X family, play important roles in regulating neuronal and glial functions in the nervous system. We have previously shown that the expression of P2X4Rs is upregulated in activated microglia after peripheral nerve injury and that activation of the receptors by extracellular ATP is crucial for maintaining nerve injury-induced pain hypersensitivity. However, the regulation of P2X4R expression on the cell surface of microglia is poorly understood. Here, we identify the CC chemokine receptor CCR2 as a regulator of P2X4R trafficking to the cell surface of microglia. In a quantitative cell surface biotinylation assay, we found that applying CCL2 or CCL12, endogenous ligands for ...
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A gray area at times. Have used the Scytek kit (check out website) for frozen sections, but did not use their peroxidase blocker, it ate frozens off the slide. Substituted a Dako peroxidase blocker for frozens was much friendlier. The kicker with this kit, their special blocker is excellent, and you have to use their chromogen with this kit. What really worked for us was to use their two blockers (not the peroxidase block though),a biotinylated primary in place of a secondary antibody with the Scytek kit since we did not have a purified primary on hand. We also diluted out the biotinylated primary further, so do a more extended dilution panel, this reduced our cross reaction background. This is the basis of the DAKO ARK kit, biotinylating the primary, some antibodies don biotinylate as well, a potential problem, but may work with yours, so another option - DAKO ARK kit. There are others on the market, Zymed, Innovex, Vector MOM kit, etc, all worth a try. Have heard some mixed reviews on Mouse ...
I wish to immobilise a peptide on a column. I think the easiest way to do this would be to biotinylate the peptide at the C-terminus and link it to streptavidin-agarose. a) Is this true? b) What is the easiest way to biotinylate specifically on the C-terminus of a peptide - do I link through a cysteine? c) How do I get rid of free biotin from a small peptide (approx. 2kD ...
Just for the sake of completing this thread, Ive decided to do a cell surface receptor biotinylation assay. Basically, incubating cells with biotin (in cold PBS) which binds to all available primary amines on the cell surface. After 1h of incubation, un-bound biotin will be inactivated by incubating the cells in 50mM Tris pH 8 for 5 min. After that, I will lyse the cells and precipitate surface proteins with streptavidin agarose. Finally, the cells surface receptor will be visualized through western blotting.. ...
Epigenetic enzymatic assays are optimized using a biotinylated histone H3-derived peptide as substrate. The modified peptide is captured by the Eu-labeled antibody (Ab) and ULight-Streptavidin (SA) which bring the Eu donor and ULight acceptor dye molecules into close proximity. Upon irradiation at 320 or 340 nm, the energy from the Eu donor is transferred to the ULight acceptor dye which, in turn, generates light at 665 nm. The intensity of the light emission is proportional to the level of biotinylated substrate modification.. ...
Epigenetic enzymatic assays are optimized using a biotinylated histone H3-derived peptide as substrate. The modified peptide is captured by the Eu-labeled antibody (Ab) and ULight-Streptavidin (SA) which bring the Eu donor and ULight acceptor dye molecules into close proximity. Upon irradiation at 320 or 340 nm, the energy from the Eu donor is transferred to the ULight acceptor dye which, in turn, generates light at 665 nm. The intensity of the light emission is proportional to the level of biotinylated substrate modification.. ...
Epigenetic enzymatic assays are optimized using a biotinylated histone H3-derived peptide as substrate. The modified peptide is captured by the Eu-labeled antibody (Ab) and ULight-Streptavidin (SA) which bring the Eu donor and ULight acceptor dye molecules into close proximity. Upon irradiation at 320 or 340 nm, the energy from the Eu donor is transferred to the ULight acceptor dye which, in turn, generates light at 665 nm. The intensity of the light emission is proportional to the level of biotinylated substrate modification.. ...
Antibodies , Primary Antibodies , Protein Tag , Anti-GST Tag, Biotinylated; Host: mouse Monoclonal; Species Reactivity: all species; Application: WB, IHC, IP
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Whatever the assay you have in mind, you need the right technologies for your target. We can custom conjugate your antibody or biomolecule to any bead, or biotinylate or DIG-label your targets, so you can develop the assay you need. If you prefer, we can even perform the assay development for you.
TY - JOUR. T1 - Ultrasensitive immuno-detection using viral nanoparticles with modular assembly using genetically-directed biotinylation. AU - Litvinov, Julia. AU - Hagström, Anna E V. AU - Lopez, Yubitza. AU - Adhikari, Meenu. AU - Kourentzi, Katerina. AU - Strych, Ulrich. AU - Monzon, Federico A.. AU - Foster, William. AU - Cagle, Philip T.. AU - Willson, Richard C.. PY - 2014. Y1 - 2014. N2 - We report a novel, modular approach to immuno-detection based on antibody recognition and PCR read-out that employs antibody-conjugated bacteriophage and easily-manipulated non-pathogenic viruses as affinity agents. Our platform employs phage genetically tagged for in vivo biotinylation during phage maturation that can easily be linked, through avidin, to any biotinylated affinity agent, including full-length antibodies, peptides, lectins or aptamers. The presence of analyte is reported with high sensitivity through real-time PCR. This approach avoids the need to clone antibody-encoding DNA fragments, ...
The attachment of biotin to various chemical sites, called biotinylation, can be used as an important laboratory technique to study various processes including DNA transcription and replication. Biotin itself is known to biotinylate histones, but is not found naturally on DNA.. Biotin binds very tightly to the tetrameric protein streptavidin, with a dissociation constant Kd in the order of 10-15 mol/L (Bonjour 1977, Green 1975) or 4x10-14 (Holmberg et al. 2005). Holmberg et al. (2005) note that the biotin-streptavidin system is the strongest noncovalent biological interaction known. This is often used in different biotechnological applications. Holmberg et al. showed how to utilize high temperatures to efficiently break the interaction without denaturation of the streptavidin.. In the biology laboratory, biotin is sometimes chemically linked, or tagged, to a molecule or protein for biochemical assays. The specificity of the biotin-streptavidin linkage allow use in molecular, immunological, and ...
If you want to get the best signal to background ratio you should go for biotinylation of your protein so you can do stringent washes. Tagging your protein with biotin ligase recognition peptide and coepressing with biotin ligase will result in the biotinylation of your protein. You can get a very good signal to background ratio this way ...
MHC multimers are oligomeric forms of MHC molecules, designed to identify and isolate T-cells with high affinity to specific antigens amid a large group of unrelated T-cells. Multimers generally range in size from dimers to octamers; however, some companies use even higher quantities of MHC per multimer. Multimers may be used to display class 1 MHC, class 2 MHC, or nonclassical molecules (e.g. CD1d) from species such as monkeys, mice, and humans. Since T-cell receptors have a low affinity for their MHC counterparts, it was historically problematic to label T cells effectively using single MHC-T-cell interactions. However, in 1996 it was proposed by John Altman to use a complex of multiple MHC molecules to form a more stable bond between corresponding T-cells. The most commonly used MHC multimers are tetramers. These are typically produced by biotinylating soluble MHC monomers, which are typically produced recombinantly in eukaryotic or bacterial cells. These monomers then bind to a backbone, ...
Bachem offers H-6098 Biotinyl-S6 Phosphate Acceptor Peptide for your research. Find all specific details here. Find product specific information including available pack sizes, CAS, detailed description and references here.
Biotin protein ligase of is the 35. or that the presence of the reaction intermediate impedes access of subtilisin to the cleavage site. The disordered loop containing residues 115C123 lies close to the biotin binding site, and residues 115C118 become ordered in the crystal structure when biotinyl-lysine occupies the active site. Because of the similarity to nucleotide-binding sequences in protein buy 118876-58-7 kinases, the sequence 115GRGRRG120 within this loop had been thought to function in ATP binding (Buoncristiani et al. 1986; Wilson et al. 1992). However, a recent mutational analysis shows that this sequence has a role in biotin binding (Kwon and Beckett 2000). No function has been identified previously for the C-terminal domain comprising residues 274C321, which shows structural similarity to the Src homology 3 domains (Noble et al. 1993). However, recent evidence of safety by biotinyl-5-AMP buy 118876-58-7 against hydroxyl radical cleavage from the BirA backbone at a number of sites ...
Adenoviral vectors have great potential for use in gene therapy and genetic immunization. The targeting of Ad vectors to the relevant tissue and cell types in vivo could greatly improve their safety and performance by lowering the effective dosage required for therapeutic levels of gene expression. Redirection of Ad vector tropism will require physical modifications of the adenoviral capsid but direct genetic modification of the Ad capsid has so far been limited to small peptides. A novel system for the attachment of targeting ligands to the Ad capsid, based on the extremely strong avidin-biotin interaction, is described herein. The genetic insertion of a biotin acceptor peptide (BAP) into the fiber, protein IX, or hexon components of the Ad capsid has resulted in vectors that are metabolically biotinylated upon production in host cells. Avidin-dependent redirection of transduction through a variety of biotinylated ligands is greatly dependent on the nature of the biotinylated capsid protein. ...
In 2012, Roux et al. published a nice paper, that received no less than four article recommendations from F1000 researchers. The paper described a method for tracking the interaction partners a protein has had within a cell (a history of its interacting partners). The method, called BioID, is based on proximity-dependent biotinylation of proteins by a promiscuous biotin ligase mutant BirA (R118G), which is fused to your protein of interest. After an overnight incubation with biotin, cells can be subjected to harsh lysis and biotinylated proteins can be isolated and identified by mass spectroscopy to determine the proteins that had come into contact with the chimeric BirA (R118G) protein. This method is a bit different from standard co-IP or pull-down experiments, because it allows one to identify proteins who interact transiently or weakly with the protein of interest. Also, due to the strong biotin-avidin binding affinity, harsh washes can greatly reduce background protein binding. [Read ...
BirA biotin ligase was expressed as maltose-binding protein (MBP) fusion in an E. coli cells. The dual 6xHis-MBP tag at the N-terminus is useful for removal of BirA after biotinylation through either Ni-NTA or amylose resin columns. Catalog number: MBBL-301 Expression host: Escherichia coli (E. coli). Expressed Region
Avidin and streptavidin reagents are powerful tools to detect or purify biotinylated proteins, nucleic acids, and other macromolecules.
1A6X: Structure of the carboxy-terminal fragment of the apo-biotin carboxyl carrier subunit of Escherichia coli acetyl-CoA carboxylase.
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In biochemistry, biotinylation is the entire process of covalently attaching biotin to a protein, nucleic acid or other molecule. Biotinylation is quick, certain and is not likely to perturb the natural operate from the molecule due to the small dimension of biotin (MW = 244.31 g/mol). Biotin binds to streptavidin and avidin with an especially superior affinity, quick on-price, and higher specificity, and these interactions are exploited in many areas of biotechnology to isolate biotinylated molecules of desire. Biotin-binding to streptavidin and avidin is resistant to extremes of heat, pH and proteolysis, earning capture of biotinylated molecules attainable in numerous types of environments ...
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Page contains details about biotin-polyA-tailed DNA-capped citrate-stabilized gold nanoparticles-coated streptavidin-coated magnetic beads . It has composition images, properties, Characterization methods, synthesis, applications and reference articles : nano.nature.com
Proven, cost-efficient LANCE Ultra reagents can be used to quantitate peptide modifications, detecting specific meythylation and acetylation states. No wash, homogenous LANCE Ultra reagents are HTS friendly - design your own epigenetics screening strategy for greatest efficiency.. Epigenetic enzymatic assays are optimized using a biotinylated histone H3-derived peptide as substrate. The modified peptide is captured by the Eu-labeled antibody (Ab) and ULight-Streptavidin (SA) which bring the Eu donor and ULight acceptor dye molecules into close proximity. Upon irradiation at 320 or 340 nm, the energy from the Eu donor is transferred to the ULight acceptor dye which, in turn, generates light at 665 nm. The intensity of the light emission is proportional to the level of biotinylated substrate modification.. ...
To determine selectivity against αIIbβIIIa, wells of a 96-well Immulon-2 microtiter plate were coated with 10 μg/ml RGD affinity-purified human αIIbβIIIa in 10 mM HEPES, 150 mM NaCl, and 1 mM MgCl2, pH 7.4 (500 ng/well αIIbβIIIa final), and then the pate was incubated overnight at 4°C. The next day, the wells were blocked with 5% BSA in the above-mentioned buffer at room temperature for 2 h. The assay plate was washed five times with modified Tyrodes buffer (150 mM NaCl, 12 mM NaHCO3, 2.6 mM KCl, 2.5 mM HEPES, 1 mM MgCl2, and 1 mg/ml BSA, pH 7.4). Biotinylated fibrinogen was prepared using human fibrinogen according to directions from the biotin-X-NHS biotinylation kit. Fifty microliters of the compound/biotinylated fibrinogen mix was transferred to the assay plate and incubated at room temperature for 2 h. After incubation, the assay plate was washed five times with modified Tyrodes buffer. Reactions were visualized as described above. JNJ-26076713 was tested at half-log doses from ...
4GJV: A dual anchoring strategy for the localization and activation of artificial metalloenzymes based on the biotin-streptavidin technology.
Purified Recombinant Human ITGAV & ITGB6 Protein, His-Avi-tagged, Biotinylated from Creative Biomart. Recombinant Human ITGAV & ITGB6 Protein, His-Avi-tagged, Biotinylated can be used for research.
SE arises from deficits in the efficacy of GABAergic inhibition that include altered GABAAR functional expression and pharmacology; however, the mechanisms underlying these deficits remain to be established (Lowenstein and Alldredge, 1998; Alldredge and Lowenstein, 1999). To address this critical question, we have examined the impact of SE on GABAAR functional expression together with the efficacy of synaptic inhibition in hippocampal slices derived from mice injected with pilocarpine. This animal model exhibits many similarities to patients undergoing SE including the development of pharmacoresistance (Kapur and Coulter, 1995; Coulter, 2001).. As measured using biotinylation, the cell surface stability of the GABAAR α1, 2, 4, β3, and γ2 subunits was significantly reduced in SE compared with controls, whereas those encoding the GABABR R1 and R2 subunits were unaltered. In contrast, the cell surface stability of the α5 and δ subunits was significantly increased. Thus, our results demonstrate ...
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Choose from among purified avidin & streptavidin proteins conjugated with a range of fluorophores for fluorescence detection of biotinylated macromolecules in a range of research applications. Click here for more.
MojoSort™ Streptavidin Nanobeads - Streptavidin Nanobeads can be used for positive or negative selection of targeted cells with biotin-conjugated antibodies.
Complete information for NRDC gene (Protein Coding), Nardilysin Convertase, including: function, proteins, disorders, pathways, orthologs, and expression. GeneCards - The Human Gene Compendium
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Sensor performance of a dielectric filled silicon bulk acoustic resonator type label-free biosensor is verified with biotin-streptavidin binding interactions as a model system. The mass sensor is a micromachined silicon square plate with a dielectric filled capacitive excitation mechanism. The resonance frequency of the biotin modified resonator decreased 315 ppm when exposed to streptavidin solution for 15 min with a concentration of 10−7 M, corresponding to an added mass of 3.43 ng on the resonator surface. An additional control is added by exposing a bovine serum albumin (BSA)-covered device to streptavidin in the absence of the attached biotin. No resonance frequency shift was observed in the control experiment, which confirms the specificity of the detection. The sensor-to-sensor variability is also measured to be 4.3%. Consequently, the developed sensor can be used to observe in biotin-streptavidin interaction without the use of labelling or molecular tags. In addition, biosensor can be used
Sensor performance of a dielectric filled silicon bulk acoustic resonator type label-free biosensor is verified with biotin-streptavidin binding interactions as a model system. The mass sensor is a micromachined silicon square plate with a dielectric filled capacitive excitation mechanism. The resonance frequency of the biotin modified resonator decreased 315 ppm when exposed to streptavidin solution for 15 min with a concentration of 10−7 M, corresponding to an added mass of 3.43 ng on the resonator surface. An additional control is added by exposing a bovine serum albumin (BSA)-covered device to streptavidin in the absence of the attached biotin. No resonance frequency shift was observed in the control experiment, which confirms the specificity of the detection. The sensor-to-sensor variability is also measured to be 4.3%. Consequently, the developed sensor can be used to observe in biotin-streptavidin interaction without the use of labelling or molecular tags. In addition, biosensor can be used
TY - JOUR. T1 - Epigenetic regulation of chromatin structure and gene function by biotin. AU - Hassan, Yousef I.. AU - Zempleni, Janos. PY - 2006/7/6. Y1 - 2006/7/6. N2 - Covalent modifications of histones are a crucial component of epigenetic events that regulate chromatin structures and gene function. Evidence exists that distinct lysine residues in histones are modified by covalent attachment of the vitamin biotin, catalyzed by biotinidase and holocarboxylase synthetase. Biotinylation of histones appears to be conserved across species. The following biotinylation sites were identified using both MS and enzymatic biotinylation of synthetic peptides: K9, K13, K125, K127, and K129 in histone H2A; K4, K9, and K18 in histone H3; and K8 and K12 in histone H4. Evidence was provided that biotinylated histone H4 is enriched in pericentromeric heterochromatin, and that biotinylation of histone H4 participates in gene silencing, mitotic condensation of chromatin, and the cellular response to DNA damage. ...
A-Kinase anchoring protein 150 (AKAP150) is required for the phosphorylation of transient receptor potential cation channel subfamily V member 1 (TRPV1) by PKA or PKC in sensory neurons and, hence, affects TRPV1-dependent hyperalgesia under pathological conditions. Recently, we showed that the activation of N-methyl-D-aspartate (NMDA) receptors sensitizes TRPV1 by enhancing serine phosphorylation through PKC in trigeminal nociceptors. In this study, we extended this observation by investigating whether AKAP150 mediates NMDA-induced phosphorylation of TRPV1 via PKC in native sensory neurons in the rat. By adopting a phospho-specific antibody combined with a surface biotinylation assay, we first assessed NMDA-induced changes in the phosphorylation level of serine 800 residues (S800) in TRPV1 delimited to cell surface membrane in cultured trigeminal ganglia (TG). The biotinylation assay yielded that the application of NMDA significantly increased the phosphorylation of S800 (p-S800) of TRPV1 at ...
U.S., Feb. 27 -- ClinicalTrials.gov registry received information related to the study (NCT03034707) titled Interference of Biotin Supplementation in Biotin-streptavidin Platforms for Hormone Testing on Jan. 17. Brief Summary: The B vitamin biotin is widely available as an over the counter supplement, often advertised and used to promote health of hair, skin and nails. Commercially available over the counter biotin supplements contain dose ranges up to 10 mg/day (ie 333 times higher than the recommended dietary allowance). The biotin molecule is also sometimes used as part of the lab technology to measure hormone and protein levels in the blood. It is possible that high doses of ingested biotin may interfere with accurate hormone or protein measurement using biotin related in vitro measurement systems. Such interference, if present, could lead to misdiagnosis. The study will analyze laboratory levels obtained with streptavidin-biotin assay systems while ingesting biotin in currently available ...
HLA-G is a nonclassical class I MHC molecule of unknown function expressed on human invasive trophoblast. In trophoblast cells, HLA-G mRNA is alternatively spliced into a variety of forms which are predicted to encode a full length membrane-bound form, three short membrane-bound isoforms and two soluble isoforms. The aim of this study was to determine which of these protein isoforms are translated, which are expressed on the cell surface and which are secreted. Artificial cDNAs encoding the isoforms were generated by PCR mutagenesis, ligated to an epitope tag and transfected into a human cell line capable of expressing MHC class I. Protein products of appropriate sizes were detected in cells transfected with cDNAs encoding all membrane-bound forms, but surface biotinylation studies indicated that only full length membrane-bound HLA-G was present at the cell surface. Full length HLA-G was also detected by surface antibody binding and flow cytometry. Soluble HLA-G1 was detected in cells transfected with
Proteins S-nitrosation represents a recently described form of post-translational modification that is rapid and reversible. in control and NO-treated cells. We recognized vimentin ubiquitin-conjugating enzyme E2 peroxiredoxin 1 β-actin and GAPDH among these proteins by a combination of two-dimensional gel electrophoresis of the biotinylated proteins and LC-MS-MS as shown in Fig. 5and Table 1. The pIs and molecular masses of these proteins highlighted in the two-dimensional gel are internally consistent (Table 1). These results were further confirmed by Western blotting showing that Tipifarnib S-nitrosation of vimentin and β-actin increased markedly after NO treatment. GAPDH which migrates as a 37.3-kDa protein band in the SDS gel as detected by both fluorescence imaging and silver staining is the major S-nitrosoprotein recognized in untreated endothelial cells. Interestingly unlike other proteins S-nitrosation of GAPDH decreased rather than increased when cells were treated with the exogenous ...
The pathological importance of tumor necrosis factor (TNF)-alpha in rheumatoid arthritis (RA) is now widely accepted. Ex vivo data from synovial cell cultures suggest that direct cell contact between activated T-cells and macrophages may be an important driver of macrophage TNF-alpha production in the RA joint. However, the ligand/receptor pairs driving this cell contact signal remain obscure. One reason for this is that plasma membrane (PM) proteins are resistant to systematic analysis using traditional proteomic approaches. In this chapter we present a method for the enrichment and resolution of PM proteins from murine T-cell hybridomas as a prelude to identification by tandem mass spectrometry. We used cell surface biotinylation, differential centrifugation and subsequent streptavidin affinity capture, followed by solution phase iso-electric focussing and tandem mass spectrometry to identify 75 PM proteins and make semiquantitative comparisons of resting and activated cells. The method is applicable
Labelling antibodies and proteins is easier than ever with SureLINK Protein Labelling Kits and Reagents! These fast, convenient, ready-to-use kits are sure to work every time whether you are labelling your proteins with biotin, enzymes, or fluroescein.. SureLINK AP Conjugation Kits and Reagents. These kits are based on novel conjugation chemistry far superior to conventional methods such as maleimide/thio, glutaraldehyde and avidin/biotin. Kits and modified enzymes are capable of conjugating a variety of proteins in a range of sizes.. SureLINK HRP Conjugation Kits and Reagents. These kits are based on the well-established periodate chemistry that yields consistent, reproducible antibody or protein conjugates. Horseradish peroxidase (HRP) is pre-activated and supplied in reaction size vials. Kits and enzymes are capable of conjugating a variety of proteins in a range of sizes.. SureLINK Chromophoric Biotin Labelling Kits. These kits provide an easy-to-use method for biotinylating antibodies and ...
Eukaryotes have evolved mechanisms to sequester mRNAs and their associated RNA-binding proteins into non-membrane delimited bodies called RNA granules as a mechanism to control translation and turnover of mRNAs. We recently identified mitochondrial RNA granules, and showed that they contain newly synthesized mitochondrial RNA, a large toolbox of proteins dedicated to RNA metabolism and the biogenesis of mitochondrial ribosomes. Using a proximity biotinylation assay (BioID) we identified a protein module containing three uncharacterized pseudouridine synthases, identified the molecular targets of these enzymes by RNA pseudoSeq, and showed that they have essential roles in epitranscriptomic modification of mitochondrial rRNA and mRNA ...
A water-soluble, enzyme co-factor present in minute amounts in every living cell. It occurs mainly bound to proteins or polypeptides and is abundant in liver, kidney, pancreas, yeast, and milk. Present in many foods; particularly rich sources include yeast, eggs, liver, certain fish (e.g. mackerel, salmon, sardines), soybeans, cauliflower and cow peas. Dietary supplement. Isol. from various higher plant sources, e.g. sweet corn seedlings and radish leaves Biotin D(+) is a cofactor responsible for carbon dioxide transfer in several carboxylase enzymes:; Biotin binds very tightly to the tetrameric protein avidin (also streptavidin and neutravidin), with a dissociation constant Kd in the order of 10?15 mol/L which is one of the strongest known protein-ligand interactions, approaching the covalent bond in strength. This is often used in different biotechnological applications. Until 2005, very harsh conditions were required to break the biotin-streptavidin bond.; Biotin is a water-soluble B-complex ...
Fluorescent Dyes , Biotins and Streptavidins , Streptavidin, recombinant; Streptavidin is a nonglycosylated, tetrameric protein, with each subunit able to bind a single molecule of the vitamin biotin. Streptavidin-biotin bond is the strongest known non-covalent interaction with Kd ~10-15 M. Because streptavidin lacks any carbohydrate modification and has a near-neutral pI, it has the advantage of much lower nonspecific binding than avidin. Streptavidin is broadly used in various applications such as immunoassays, histochemistry, FISH (Fluorescence In Situ Hybridization), flow cytometry, microarrays and blot analysis.
General protocols for optimizing and conducting surface biotinylation labeling assays have been described previously (Gottardi and Caplan 1992; Gottardi et al. 1995). Briefly, 1°PVD epithelial cells derived from each of four boars were cultured as confluent monolayers on permeable supports in two 6‐well Transwell plates (12 inserts, 4.67 cm2 per insert). All monolayers were washed free of serum proteins in three changes of Ringer solution (composition in mmol/L: 120 NaCl, 25 NaHCO3, 3.3 KH2PO4, 0.83 K2HPO4, 1.2 CaCl2, 1.2 MgCl2) and then incubated in Ringer solution at 37°C for 30 min. Monolayers were gradually cooled to ~4°C on ice for 30 min followed by exposure to vehicle or biotin (EZ‐link® Sulfo‐NHS‐LC‐Biotin, Pierce, Rockford, IL) either apically or basolaterally (three wells, each) for 30 min with gentle agitation. At the end of the incubation period, apical and basolateral solutions were removed and cells were washed five times (3 min each) with 50 mmol/L glycine in Ringer ...
Streptavidin is a non-glycosylated protein originally isolated from bacterium Streptomyces avidinii. With a very high affinity for biotin, it is widely used to bridge biotinylated probes and enzymes.
Streptavidin (SA) is produced by Streptomyces avidinii. SA has an extraordinarily high affinity for biotin with the advantage of much lower NBS than avidin
The coefficient of variation from well to well is under 5 % for 96 well microplates and under 8 % for 384 well microplates. The streptavidin solid phase is treated with an additional blocking step in order to minimise any unspecific binding, therefore, „pre-blocking" of plates is not necessary. The high stability of the coating and the high affinity between streptavidin and biotin enables unusually stringent washing conditions, which have ...
Read independent reviews on Biotinylated hyaluronic acid (hyaluronan) plate from AMS Biotechnology (Archived Products) on SelectScience
Biotinylated synaptophysin does not appear in SLMVs at 18°C. PC12 cells were incubated with sulfoNHS-LC-biotin either for 30 min at 37°C (A), for 30 min
gp120 binding by and competition of R880F mAbs 19.3H-L1 and 19.3H-L3.The baseline binding of four biotinylated mAbs, 19.3H-L1, 19.3H-L3, 6.4C, or PGT128, was ev
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Holocarboxylase synthetase (HCS, human) and BirA (Escherichia coli) are biotin protein ligases that catalyze the ATP-dependent attachment of biotin to apocarboxylases. Biotin attachment occurs on a highly conserved lysine residue within a consensus sequence (Ala/Val-Met-Lys-Met) that is found in carboxylases in most organisms. Numerous studies have indicated that HCS and BirA, as well as biotin protein ligases from other organisms, can attach biotin to apocarboxylases from different organisms, indicating that the mechanism of biotin attachment is well conserved. In this study, we examined the cross-reactivity of biotin attachment between human and bacterial biotin ligases by comparing biotinylation of p-67 and BCCP87, the biotin-attachment domain fragments from human propionyl-CoA carboxylase and E. coli acetyl-CoA carboxylase, respectively. While BirA has similar biotinylation activity toward the two substrates, HCS has reduced activity toward bacterial BCCP87 relative to its native substrate, p-67.
Biotin is required for enzymatic activity of several important carboxylases (Table 28.10). Biotin is first attached to a lysine residue of biotin carboxyl carrier protein (BCCP) by biotin- [propionyl-CoA-carboxylase (ATP-hydrolysing)] ligase (holocarboxylase [HCS], EC 6.3.4.10), which forms the biotinyl domain of the enzyme or separate subunit. The N1 position is subsequently carboxylated to form carboxy-biotin (Fig. 28.40), which is then able to transfer the carboxyl group to other substrates. The source of the carboxyl is either bicarbonate or decarboxylation of another substrate as is seen in transcarboxylation reactions.196 A typical carboxylation reaction mediated by biotin-BCCP is shown in Figure 28.41. The source of the carboxyl is bicarbonate with ATP supplying energy for the reaction. This is followed by the subsequent transfer of the carboxyl to the substrate with regeneration of the biotin-BCCP complex.. An interesting recent development is the finding that biotin is also attached to ...
Mycobacterium tuberculosis (Mtb), a dreaded pathogen, has a unique cell envelope composed of high fatty acid content that plays a crucial role in its pathogenesis. Acetyl Coenzyme A Carboxylase (ACC), an important enzyme that catalyzes the first reaction of fatty acid biosynthesis, is biotinylated by biotin acetyl-CoA carboxylase ligase (BirA). The ligand-binding loops in all known apo BirAs to date are disordered and attain an ordered structure only after undergoing a conformational change upon ligand-binding. Here, we report that dehydration of Mtb-BirA crystals traps both the apo and active conformations in its asymmetric unit, and for the first time provides structural evidence of such transformation. Recombinant Mtb-BirA was crystallized at room temperature, and diffraction data was collected at 295 K as well as at 120 K. Transfer of crystals to paraffin and paratone-N oil (cryoprotectants) prior to flash-freezing induced lattice shrinkage and enhancement in the resolution of the X-ray ...
Video articles in JoVE about bacteriophage t7 include Kinetics of Lagging-strand DNA Synthesis In Vitro by the Bacteriophage T7 Replication Proteins, Protocols for Implementing an Escherichia coli Based TX-TL Cell-Free Expression System for Synthetic Biology, Synthesis of Infectious Bacteriophages in an E. coli-based Cell-free Expression System, Rescue of Recombinant Newcastle Disease Virus from cDNA, Expression, Isolation, and Purification of Soluble and Insoluble Biotinylated Proteins for Nerve Tissue Regeneration, Reverse Genetics Mediated Recovery of Infectious Murine Norovirus, Whole Mount RNA Fluorescent in situ Hybridization of Drosophila Embryos, Direct Observation of Enzymes Replicating DNA Using a Single-molecule DNA Stretching Assay, Nuclear Magnetic Resonance Spectroscopy for the Identification of Multiple Phosphorylations of Intrinsically Disordered Proteins, Using Tomoauto: A Protocol for High-throughput Automated Cryo-electron Tomography, Visualizing Single-molecule
Gap junctions are specialized cell-cell contacts that provide direct intercellular communication between eukaryotic cells. The tyrosine-sorting signal (YXXØ), present at amino acids 286-289 of Cx43 (connexin43), has been implicated in the internalization of the protein. In recent years, ubiquitination of Cx43 has also been proposed to regulate gap junction intercellular communication; however, the underlying mechanism and molecular players involved remain elusive. In the present study, we demonstrate that ubiquitinated Cx43 is internalized through a mechanism that is independent of the YXXØ signal. Indeed, expression of a Cx43-Ub (ubiquitin) chimaera was shown to drive the internalization of a mutant Cx43 in which the YXXØ motif was eliminated. Immunofluorescence, cycloheximide-chase and cell-surface-protein biotinylation experiments demonstrate that oligomerization of Cx43-Ub into hemichannels containing wild-type Cx43 or mutant Cx43Y286A is sufficient to drive the internalization of the ...
In live cells, fluorescent E-cadherin was internalized into macropinosomes from non-contact cell edges, but then it was often seen being recycled to regions of cell-cell contact. Macropinocytosis might then also be a mechanism for redeploying E-cadherin on the cell surface to modify regions of cell-cell contact in response to growth factors. Macropinocytosis and recycling of other adhesion proteins, such as the tight-junction proteins occludin and JAM-A occurs in response to IFNγ stimulation without protein downregulation, to facilitate temporary trans-epithelial permeability (Bruewer et al., 2005; Utech et al., 2005). Our results here show too that EGF induces trafficking of E-cadherin without overt loss of the total protein levels. However, biotinylation experiments revealed that there is an increase in the turnover of E-cadherin in cells silenced for SNX1 expression, suggesting a role for SNX1 in sorting (for either recycling or degradation) of endocytic membrane containing internalized ...
One of the most popular methods of noncovalent conjugation is to make use of the natural strong binding of avidin or its derivative streptavidin to biotin. Each avidin molecule contains a maximum of four biotin binding sites thereby increasing the strength of their interaction with biotin. Depending on the functionality present on the biotinylation compounds, specific reactive groups on antibodies may be modified to create a avidin binding site. Amines, carboxylates, sulfhydryls, and carbohydrate groups can be specifically targeted for biotinylation through the appropriate choice of biotin derivative.. Avidin is a glycoprotein found in egg whites that contains four identical subunits of 16,400 Da each. The subunits each contain one binding site for biotin also known as vitamin H. The biotin interaction with avidin is the strongest noncovalent affinity known, exhibiting a dissociation constant of about 1.3×10-15M. Tryptophan and lysine residues in each subunit are involved in forming the binding ...
... : A Coggle Diagram about Chapter 2 (Result 1 Hematological cancer cell line screen, HAI-2 complex validation, Activity validation, HAI-2/MTP ratio validation, HAI-2 gradient expression, HAI-1 gradient expression, Leakage expression and activity, Surface biotinylation depletion, IHC and Summary), B cell lymphoma (Tumor microenvironment in B cell lymphoma, Signaling pathway in B cell lymphoma, Treatment, Classification, Differentiation of B cell and Mutation), Matriptase (Substrate, Regulation, Lymphoma, Physiological function, Tumorigenic role and Trafficking) and Chapter 3
Our family of biotin-binding proteins includes streptavidin, avidin, and NeutrAvidin® protein. See all conjugated and unconjugated streptavidin products.
Victor Muñoz Robles, Marc Dürrenberger, Tillmann Heinisch, Agustí Lledós, Tilman Schirmer, Thomas R. Ward, Jean-Didier Maréchal, Structural, Kinetic, and Docking Studies of Artificial Imine Reductases Based on Biotin-Streptavidin Technology: An Induced Lock-and-Key Hypothesis, Journal of the American Chemical Society, 2014, 136, 44, ...
Purified Recombinant Human CD47 protein, Fc/Avi-tagged, Biotinylated from Creative Biomart. Recombinant Human CD47 protein, Fc/Avi-tagged, Biotinylated can be used for research.
Alongside the increasing availability of affinity reagents, antibody microarrays have been developed to become a powerful tool to screen for target proteins in complex samples. Besides multiplexed sandwich immunoassays, the application of directly applying labeled sample onto arrays with immobilized capture reagents offers an approach to facilitate a systematic, high-throughput analysis of body fluids such as serum or plasma. An alternative to commonly used planar arrays has become available in form of a system based on color-coded beads for the creation of antibody arrays in suspension. The assay procedure offers an uncomplicated option to screen larger numbers of serum or plasma samples with variable sets of capture reagents. In addition, the established procedure of whole sample biotinylation circumvents the purification steps, which are generally required to remove excess labeling substance. We have shown that this assay system allows detecting proteins down into lower pico-molar and higher ...
Animal, Animal Models, Biotin, Biotinylation, Birth, Birth Defects, Chromatin, Epigenetic, Gene, Genome, Genome Instability, Genome Stability, Histone, Histones, Human, Long Terminal Repeats, Methylation, Multiprotein Complexes, Population, Repression
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Page contains details about [email protected] modified DNA nanoribbon 100:1 . It has composition images, properties, Characterization methods, synthesis, applications and reference articles : nano.nature.com
Monoclonal Antibody for studying Akt1 (Ser473) phosphate/Akt2 (Ser474) phosphate/Akt3 (Ser472) phosphate in the PI3K / Akt Signaling research area.
NRDC has long collaborated with regional partners, including ranchers, to promote nonlethal methods as an effective alternative to the needless cycles of conflict and killing.
Known proteins associated with the cell-adhesion protein E-cadherin include catenins and proteins involved in signaling, trafficking and actin organization. However, the list of identified adherens junction proteins is likely to be incomplete, limiting investigation into this critical cell structure. To expand the inventory of potentially relevant proteins, we expressed E-cadherin fused to biotin ligase in MDCK epithelial cells, and identified by mass spectrometry neighboring proteins which were biotinylated. The most abundant of the 303 proteins identified were catenins and nearly 40 others that had been previously reported to influence cadherin function. Many others could be rationalized as novel candidates for regulating the adherens junction, cytoskeleton, trafficking or signaling. We further characterized lipoma preferred partner (LPP), which is present at both cell-contacts and focal adhesions. Knockdown of LPP demonstrated its requirement for E-cadherin dependent adhesion and suggested it ...
In the first part of this work, microfabricated Capillary Electrophoresis devices were integrated with microfabricated PCR devices for fast DNA amplification and separation., rapid on-chip amplification and separation of genes of interest was performed. A micro DNA extraction device using paramagnetic micro silica beads was consequently developed, subsequently a combined on-chip DNA extraction from a single drop of blood and PCR amplification were performed. In the second part of this work, micro array-based strategies for proteomics studies were developed. Two new strategies for site-specific immobilization of kinase substrates onto functionalized slides were developed: In addition, a novel fluorescent antibody-based detection was developed to rapidly screen for kinase substrate phosphorylation. Avidin-biotin is one of the strongest know non-covalent interaction, and by arraying C-terminal biotinylated proteins on avidin functionalized slides, a very stable array of functionally active proteins ...
K11263 bccA; acetyl-CoA/propionyl-CoA carboxylase, biotin carboxylase, biotin carboxyl carrier protein [EC:6.4.1.2 6.4.1.3 6.3.4.14 ...
HCS antibody (holocarboxylase synthetase (biotin-(proprionyl-CoA-carboxylase (ATP-hydrolysing)) ligase)) for IHC-P, WB. Anti-HCS pAb (GTX109815) is tested in Human samples. 100% Ab-Assurance.
... (Gold NanoUrchin Protein Conjugates). Streptavidin conjugated 100nm gold nanourchins (0.5 ml, OD10)
Kazi, J.A., Liu, E.H.C., Lee, T.L., Tachibana, S. (2007). Localization of nocistatin-binding sites in mice brain and spinal cord using a biotinylated nocistatin probe. NeuroReport 18 (8) : 767-770. [email protected] Repository. https://doi.org/10.1097/WNR. ...
Read independent reviews on Covalent coated Streptavidin glass slide (5 Slides/Box) from AutoMate Scientific Inc. on SelectScience
ForteBios Super Streptavidin biosensors offer the highest sensitivity for the demands of small molecule kinetic experiments. For Analysis of Small Molecule-Protein Interactions and Fragment Screening
... ,Applications include ELISA, immunochemistry, and Western blotting. Working Concentrations: ELISA: 0.005-0.05 U/ml, immunohisto/cytochemistry: 0.25-0.5 U/ml, Western blot: 0.1-0.25 U/ml.,biological,biology supply,biology supplies,biology product
Bifunctional ligase/repressor BirA; Acts both as a biotin--[acetyl-CoA-carboxylase] ligase and a biotin-operon repressor. In the presence of ATP, BirA activates biotin to form the BirA-biotinyl-5-adenylate (BirA-bio- 5-AMP or holoBirA) complex. HoloBirA can either transfer the biotinyl moiety to the biotin carboxyl carrier protein (BCCP) subunit of acetyl-CoA carboxylase, or bind to the biotin operator site and inhibit transcription of the operon (319 aa ...
Since favorable images of infection are obtained with radio-labeled nonspecific IgG, streptavidin has been considered as an alternative protein in this investigation. The advantage of streptavidin is that once localized it may be targeted with radiolabeled biotin. Studies were conducted in a mouse model with an Escherichia coli infection in one thigh. Indium-111-labeled streptavidin showed equivalent localization to the infection as that obtained with 111In-labeled polyclonal nonspecific IgG, however blood levels with streptavidin were lower at all time points; consequently, target-to-blood ratios were improved. Pretargeting with unlabeled streptavidin followed 3 hr later with 111In-labeled biotin showed equivalent localization in the target and reduced activity in all organs sampled. As such, infected thigh-to-normal thigh ratios were improved 3-fold for pretargeting versus either labeled IgG or streptavidin. Improvements in infected thigh-to-liver and blood ratios were greater than 8-fold. Only in the
We used domain-selective biotinylation/125I-streptavidin blotting (Sargiacomo, M., M. P. Lisanti, L. Graeve, A. Le Bivic, and E. Rodriguez-Boulan. 1989 J. Membr. Biol. 107:277-286), in combination with lectin precipitation, to analyze the apical and basolateral glycoprotein composition of Madin-Darby canine kidney (MDCK) cells and to explore the role of glycosylation in the targeting of membrane glycoproteins. All six lectins used recognized both apical and basolateral glycoproteins, indicating that none of the sugar moieties detected were characteristic of the particular epithelial cell surface. Pulse-chase experiments coupled with domain-selective glycoprotein recovery were designed to detect the initial appearance of newly synthesized glycoproteins at the apical or basolateral cell surface. After a short pulse with a radioactive precursor, glycoproteins reaching each surface were biotinylated, extracted, and recovered via precipitation with immobilized streptavidin. Several basolateral ...
biotinylation: covalent attachment of a biotin moiety using a biotinylation reagent, typically for the purpose of labeling a ...
This biotinylation reaction requires ATP and is catalyzed by holocarboxylase synthetase. In bacteria, biotin is attached to ... This process is called biotinylation. Because both streptavidin and avidin bind biotin with high affinity (Kd of 10−14 mol/l to ... Biotinylation Multiple carboxylase deficiency NeutrAvidin Photobiotin Strep-tag Streptavidin Merck Index, 11th Edition, 1244. " ... Hymes J, Fleischhauer K, Wolf B (1995). "Biotinylation of histones by human serum biotinidase: assessment of biotinyl- ...
... biotinylation or glucose decomposition. Typically, the primary building blocks of these nanostructures are individual ...
Biotinylation of DNA and RNA with photoactivatable biotin is easier and less expensive than enzymatic methods since the DNA and ...
"Identification of new accessible tumor antigens in human colon cancer by ex vivo protein biotinylation and comparative mass ...
Chapman-Smith A, Cronan JE (September 1999). "The enzymatic biotinylation of proteins: a post-translational modification of ...
NHS-coupling gives biotinylation of any primary amines in the protein). Enzymatic biotinylation results in biotinylation of a ... Chemical biotinylation utilises various conjugation chemistries to yield nonspecific biotinylation of amines, carboxylates, ... In contrast to chemical biotinylation methods, enzymatic biotinylation allows biotin to be linked at exactly one residue ... Photoactivatable biotinylation reagents can also be used to activate biotinylation at specific times in an experiment or during ...
Eggnog Meringue Mousse Smoothie Streptavidin Biotinylation Nurminen et al. 2007 Green 1963 Korpela 1984 Green 1975 Bruch & ...
Limitations For Chem-seq to be feasible, the small molecule under study must be amenable to biotinylation without disruption of ...
Protein biotinylation in vivo was proposed to alleviate the problems caused by frequent incompatibility of antibody staining ... Viens, A.; Harper, F.; Pichard, E.; Comisso, M.; Pierron, G.; Ogryzko, V. (2008). "Use of Protein Biotinylation in Vivo for ...
The advent of proximity biotinylation by birA* has facilitated the first proteomics-based studies of the cadherin adhesome. ...
... such as biotinylation by biotin ligase) or chemical modification (such as reaction with FlAsH-EDT2 for fluorescence imaging). ... a peptide allowing biotinylation by the enzyme BirA and so the protein can be isolated by streptavidin (GLNDIFEAQKIEWHE) ...
Chapman-Smith A, Cronan JE (1999). "The enzymatic biotinylation of proteins: a post-translational modification of exceptional ...
One idea is to attach the exonuclease to the nanopore, perhaps through biotinylation to the beta barrel hemolysin. The central ...
Subtiligase biotinylation of N-termini uses enzymatic labeling of N-terminal peptides, but does not use lysine blocking ... Lysine guanidination followed by biotinylation of N-termini uses a chemical to block lysine residues and tag free N-termini. ... Acetylation of amines followed by tryptic digestion and biotinylation of free N-terminal peptides uses chemical (acetylation) ...
2004). "Use of protein biotinylation in vivo for chromatin immunoprecipitation". Analytical Biochemistry. 325 (1): 68-76. doi: ... or in vivo biotinylation can be used instead of antibodies to the native protein of interest. The DNA associated with the ...
... can act as an ATP sensor protein through biotinylation. Biotinylation will immobilize luciferase on the cell-surface ...
Exploiting these facts, scientists have bioengineered E. coli to produce soluble MHC molecules with a biotinylation protein ...
Biotinylation holocarboxylase synthetases at the US National Library of Medicine Medical Subject Headings (MeSH). ...
Citrulline residues can be chemically modified with butanedione or biotinylation prior to analysis, leading to a different mass ... Tutturen, Astrid E. V.; Holm, Anders; Fleckenstein, Burkhard (2013-11-01). "Specific biotinylation and sensitive enrichment of ...
... biotinylation, formylation, geranylgeranylation, glutamylation, glycosylation, glycylation, hydroxylation, isoprenylation, ...
... thereby increasing the abundance of histone H4 biotinylation marks in HEK-293 human kidney cells". The Journal of Nutritional ...
Fouda AE, Pflum MK (2015). "A Cell-Permeable ATP Analogue for Kinase-Catalyzed Biotinylation". Angew. Chem. Int. Ed. 54: 9618- ... "The generality of kinase-catalyzed biotinylation". Bioorganic & Medicinal Chemistry. 24 (1): 12-9. doi:10.1016/j.bmc.2015.11. ...
A minimal peptide substrate in biotin holoenzyme synthetase-catalyzed biotinylation. Protein Sci. 1999;8(4):921-9. http://dx. ...
While BirA has similar biotinylation activity toward the two substrates, HCS has reduced activity toward bacterial BCCP87 ... we examined the cross-reactivity of biotin attachment between human and bacterial biotin ligases by comparing biotinylation of ... While BirA has similar biotinylation activity toward the two substrates, HCS has reduced activity toward bacterial BCCP87 ... we examined the cross-reactivity of biotin attachment between human and bacterial biotin ligases by comparing biotinylation of ...
In Vivo Biotinylation in E.coli. FGFR4 Proteins for Coturnix coturnix (Common quail) (Tetrao coturnix) ...
One way is to insert a Affinity Chromatography: Principles and Applications 21 biotinylation sequence into a recombinant ...
A comprehensive platform for the analysis of ubiquitin-like protein modifications using in vivo biotinylation * L Pirone ...
Antibody Conjugation - Biotinylation. *Antibody Conjugation - FITC. *Antibody Pair for Proximity Ligation Assay ...
Sulfo-NHS-Biotin is a water soluble biotinylation reagent. Sulfo-NHS-Biotin is ideal for biotinylating antibody or protein that ... Home » Products » Biotinylation Reagents » Sulfosuccinimidyl biotin (Sulfo-NHS-Biotin). My Account , Cart Contents , Checkout ...
Versatile-removes unreacted fluorescent dyes, biotinylation reagents, crosslinkers, and reducing agents from protein solutions ... Versatile-removes unreacted fluorescent dyes, biotinylation reagents, crosslinkers, and reducing agents from protein solutions ... Versatile-removes unreacted fluorescent dyes, biotinylation reagents, crosslinkers, and reducing agents from protein solutions ... Versatile-removes unreacted fluorescent dyes, biotinylation reagents, crosslinkers, and reducing agents from protein solutions ...
Grow E. coli containing the biotinylation plasmid (with the target antigen inserted as a fusion to the N-terminus of BCCP) O/N ...
In Vivo Biotinylation in E.coli *Baculovirus. *Mammalian cell. Code. CSB-YP629626SAAU. ...
The bacteriophages have been genetically engineered to express AviTags at their ends, which permit biotinylation and their ...
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Features of our Biotinylated Kinases are: Biotinylation at a single site outside of the catalytic domain, enzymatic activity of ...
Reactive CF® Dyes, Other Reactive Dyes & Biotinylation Reagents. Antibody & Protein Labeling Kits ...
Reactive CF® Dyes, Other Reactive Dyes & Biotinylation Reagents. *Reagents for Nitric Oxide (NO) & Reactive Oxygen Species (ROS ...
1A, lane IV). In these experiments, CCR5 was not detected because of the low efficiency of biotinylation; however, it was ... Because CD4 is easier to detect by Western blotting and biotinylation than CCR5, in most cases we used anti-CCR5 antibodies to ... we used silver staining of proteins immunoprecipitated by the anti-CCR5 mAb 5C7 in parallel with biotinylation (some proteins, ...
... of transplanted cell populations with superparamagnetic iron oxide nanoparticles using cell surface chemical biotinylation for ... of transplanted cell populations with superparamagnetic iron oxide nanoparticles using cell surface chemical biotinylation for ...
Cell surface biotinylation. Increase in plasma membrane level. 14517216. TRPV4. MAP7. Patch clamp. Activation. 14517216. ...
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MATERIALS Bis-biotinylation tags Patent US9678080B2 Original Assignee Pacific Biosciences of California Inc Priority date 2013- ...
EZ-Link™ NHS-PEG4 Biotinylation Kit. EZ-Link™ Micro NHS-PEG4-Biotinylation Kit ... NHS-PEG4-Biotin is a long (29.0Å), pegylated, water-soluble, NHS-ester biotinylation reagent to label amines and maximize ... N-Hydroxysulfosuccinimide (NHS) esters of biotin are the most popular type of biotinylation reagent. NHS-activated biotins ...
E) Red blood cell half-life measured in vivo using biotinylation, showing a more rapid rate of red blood cell turnover in the ... Red blood cell half-life measured in vivo in 7-week post-transplanted mice using biotinylation, demonstrating the increased ...
AirID, a novel proximity biotinylation enzyme, for analysis of protein-protein interactions Kohki Kido et al. ... AirID provides highly interaction-dependent biotinylation for analysis of protein-protein interaction. ...
Surface biotinylation assays in CF cells confirmed the ability of the PI3Kγ to promote trafficking of the mutant CFTR to the ... chloride current measurements and surface biotinylation assays were carried out in bronchial primary CF and CFBE41o- cells. ...
Sodium Monensin, isolated from Streptomyces cinnamonensis, is a well-known representative of naturally polyether ionophore antibiotics.
  • In this study, we examined the cross-reactivity of biotin attachment between human and bacterial biotin ligases by comparing biotinylation of p-67 and BCCP87, the biotin-attachment domain fragments from human propionyl-CoA carboxylase and E. coli acetyl-CoA carboxylase, respectively. (ox.ac.uk)
  • Please note that the reagents listed on this page have been produced by the NIH Tetramer Facility with strict quality control (including gene sequencing, affinity purification, SDS PAGE analysis, and verification of biotinylation by immunoprecipitation), but many have not been confirmed to stain or activate specific T cells. (emory.edu)
  • BIC has developed a series of protein labeling techniques, including fluorescent probes (FITC), enzyme conjugates (horseradish peroxidase and alkaline phosphatase), isotope labeling and biotinylation. (biologicscorp.com)
  • The proposed single-molecule AFM approach will be instrumental for studying the effects of various epigenetic modifications of nucleosomes, in addition to biotinylation. (saladgaffe.gq)
  • Biotinylation can take place at the binding site inhibiting subsequent analyte interaction but that is not expected with the HIS-tag. (sprpages.nl)