A thermostable extracellular metalloendopeptidase containing four calcium ions. (Enzyme Nomenclature, 1992) 3.4.24.27.
ENDOPEPTIDASES which use a metal such as ZINC in the catalytic mechanism.
Peptides composed of two amino acid units.
Serum globulins with high molecular weight. (Dorland, 28th ed)
A serine endopeptidase that is formed from TRYPSINOGEN in the pancreas. It is converted into its active form by ENTEROPEPTIDASE in the small intestine. It catalyzes hydrolysis of the carboxyl group of either arginine or lysine. EC 3.4.21.4.
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.
A brominating agent that replaces hydrogen atoms in benzylic or allylic positions. It is used in the oxidation of secondary alcohols to ketones and in controlled low-energy brominations. (From Miall's Dictionary of Chemistry, 5th ed; Hawley's Condensed Chemical Dictionary, 12th ed,).
Partial proteins formed by partial hydrolysis of complete proteins or generated through PROTEIN ENGINEERING techniques.
A serine endopeptidase secreted by the pancreas as its zymogen, CHYMOTRYPSINOGEN and carried in the pancreatic juice to the duodenum where it is activated by TRYPSIN. It selectively cleaves aromatic amino acids on the carboxyl side.
A genus of BACILLACEAE that are spore-forming, rod-shaped cells. Most species are saprophytic soil forms with only a few species being pathogenic.
Compounds which inhibit or antagonize biosynthesis or actions of proteases (ENDOPEPTIDASES).
A subclass of PEPTIDE HYDROLASES that catalyze the internal cleavage of PEPTIDES or PROTEINS.
Proteins which contain carbohydrate groups attached covalently to the polypeptide chain. The protein moiety is the predominant group with the carbohydrate making up only a small percentage of the total weight.
The process of cleaving a chemical compound by the addition of a molecule of water.
Enzyme that is a major constituent of kidney brush-border membranes and is also present to a lesser degree in the brain and other tissues. It preferentially catalyzes cleavage at the amino group of hydrophobic residues of the B-chain of insulin as well as opioid peptides and other biologically active peptides. The enzyme is inhibited primarily by EDTA, phosphoramidon, and thiorphan and is reactivated by zinc. Neprilysin is identical to common acute lymphoblastic leukemia antigen (CALLA Antigen), an important marker in the diagnosis of human acute lymphocytic leukemia. There is no relationship with CALLA PLANT.
A metallic element of atomic number 30 and atomic weight 65.38. It is a necessary trace element in the diet, forming an essential part of many enzymes, and playing an important role in protein synthesis and in cell division. Zinc deficiency is associated with ANEMIA, short stature, HYPOGONADISM, impaired WOUND HEALING, and geophagia. It is known by the symbol Zn.
The extent to which an enzyme retains its structural conformation or its activity when subjected to storage, isolation, and purification or various other physical or chemical manipulations, including proteolytic enzymes and heat.
Carboxypeptidases that are primarily found the DIGESTIVE SYSTEM that catalyze the release of C-terminal amino acids. Carboxypeptidases A have little or no activity for hydrolysis of C-terminal ASPARTIC ACID; GLUTAMIC ACID; ARGININE; LYSINE; or PROLINE. This enzyme requires ZINC as a cofactor and was formerly listed as EC 3.4.2.1 and EC 3.4.12.2.
The parts of a macromolecule that directly participate in its specific combination with another molecule.
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.
Hydrolases that specifically cleave the peptide bonds found in PROTEINS and PEPTIDES. Examples of sub-subclasses for this group include EXOPEPTIDASES and ENDOPEPTIDASES.
Cyanogen bromide (CNBr). A compound used in molecular biology to digest some proteins and as a coupling reagent for phosphoroamidate or pyrophosphate internucleotide bonds in DNA duplexes.
A potent inhibitor of membrane metalloendopeptidase (ENKEPHALINASE). Thiorphan potentiates morphine-induced ANALGESIA and attenuates naloxone-precipitated withdrawal symptoms.
Organic compounds that generally contain an amino (-NH2) and a carboxyl (-COOH) group. Twenty alpha-amino acids are the subunits which are polymerized to form proteins.
The rate dynamics in chemical or physical systems.
A process of preserving animal hides by chemical treatment (using vegetable tannins, metallic sulfates, and sulfurized phenol compounds, or syntans) to make them immune to bacterial attack, and subsequent treatments with fats and greases to make them pliable. (McGraw-Hill Dictionary of Scientific and Technical Terms, 5th ed)
Methods used to remove unwanted facial and body hair.
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.
The lipid- and protein-containing, selectively permeable membrane that surrounds the cytoplasm in prokaryotic and eukaryotic cells.
The protein complement of an organism coded for by its genome.
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.
A colorless liquid used as a solvent and an antiseptic. It is one of the ketone bodies produced during ketoacidosis.
Four or five slender jointed digits in humans and primates, attached to each HAND.
Any one of five terminal digits of the vertebrate FOOT.
The meaning ascribed to the BASE SEQUENCE with respect to how it is translated into AMINO ACID SEQUENCE. The start, stop, and order of amino acids of a protein is specified by consecutive triplets of nucleotides called codons (CODON).
An iron-binding beta1-globulin that is synthesized in the LIVER and secreted into the blood. It plays a central role in the transport of IRON throughout the circulation. A variety of transferrin isoforms exist in humans, including some that are considered markers for specific disease states.
A metallic element with atomic symbol Fe, atomic number 26, and atomic weight 55.85. It is an essential constituent of HEMOGLOBINS; CYTOCHROMES; and IRON-BINDING PROTEINS. It plays a role in cellular redox reactions and in the transport of OXYGEN.
Negatively charged atoms, radicals or groups of atoms which travel to the anode or positive pole during electrolysis.
Membrane glycoproteins found in high concentrations on iron-utilizing cells. They specifically bind iron-bearing transferrin, are endocytosed with its ligand and then returned to the cell surface where transferrin without its iron is released.
Organic chemicals that form two or more coordination links with an iron ion. Once coordination has occurred, the complex formed is called a chelate. The iron-binding porphyrin group of hemoglobin is an example of a metal chelate found in biological systems.
Proteins that specifically bind to IRON.
The interchange of goods or commodities, especially on a large scale, between different countries or between populations within the same country. It includes trade (the buying, selling, or exchanging of commodities, whether wholesale or retail) and business (the purchase and sale of goods to make a profit). (From Random House Unabridged Dictionary, 2d ed, p411, p2005 & p283)
Governmental levies on property, inheritance, gifts, etc.
The charge levied on the consumer for drugs or therapy prescribed under written order of a physician or other health professional.
Study of stamps or postal markings. It usually refers to the design and commemorative aspects of the stamp.
Traditional Arabic methods used in medicine in the ARAB WORLD.
Bone-growth regulatory factors that are members of the transforming growth factor-beta superfamily of proteins. They are synthesized as large precursor molecules which are cleaved by proteolytic enzymes. The active form can consist of a dimer of two identical proteins or a heterodimer of two related bone morphogenetic proteins.
Exclusive legal rights or privileges applied to inventions, plants, etc.
A potent osteoinductive protein that plays a critical role in the differentiation of osteoprogenitor cells into OSTEOBLASTS.
A bone morphogenetic protein that is a potent inducer of bone formation. It also functions as a regulator of MESODERM formation during EMBRYONIC DEVELOPMENT.
A bone morphogenetic protein that is widely expressed during EMBRYONIC DEVELOPMENT. It is both a potent osteogenic factor and a specific regulator of nephrogenesis.
A specialized CONNECTIVE TISSUE that is the main constituent of the SKELETON. The principle cellular component of bone is comprised of OSTEOBLASTS; OSTEOCYTES; and OSTEOCLASTS, while FIBRILLAR COLLAGENS and hydroxyapatite crystals form the BONE MATRIX.
A subtype of bone morphogenetic protein receptors with high affinity for BONE MORPHOGENETIC PROTEINS. They can interact with and undergo PHOSPHORYLATION by BONE MORPHOGENETIC PROTEIN RECEPTORS, TYPE II. They signal primarily through RECEPTOR-REGULATED SMAD PROTEINS.

Different regulation of the p53 core domain activities 3'-to-5' exonuclease and sequence-specific DNA binding. (1/552)

In this study we further characterized the 3'-5' exonuclease activity intrinsic to wild-type p53. We showed that this activity, like sequence-specific DNA binding, is mediated by the p53 core domain. Truncation of the C-terminal 30 amino acids of the p53 molecule enhanced the p53 exonuclease activity by at least 10-fold, indicating that this activity, like sequence-specific DNA binding, is negatively regulated by the C-terminal basic regulatory domain of p53. However, treatments which activated sequence-specific DNA binding of p53, like binding of the monoclonal antibody PAb421, which recognizes a C-terminal epitope on p53, or a higher phosphorylation status, strongly inhibited the p53 exonuclease activity. This suggests that at least on full-length p53, sequence-specific DNA binding and exonuclease activities are subject to different and seemingly opposing regulatory mechanisms. Following up the recent discovery in our laboratory that p53 recognizes and binds with high affinity to three-stranded DNA substrates mimicking early recombination intermediates (C. Dudenhoeffer, G. Rohaly, K. Will, W. Deppert, and L. Wiesmueller, Mol. Cell. Biol. 18:5332-5342), we asked whether such substrates might be degraded by the p53 exonuclease. Addition of Mg2+ ions to the binding assay indeed started the p53 exonuclease and promoted rapid degradation of the bound, but not of the unbound, substrate, indicating that specifically recognized targets can be subjected to exonucleolytic degradation by p53 under defined conditions.  (+info)

The cyclic structure of microcin J25, a 21-residue peptide antibiotic from Escherichia coli. (2/552)

Microcin J25 (MccJ25) is the single representative of the immunity group J of the microcin group of peptide antibiotics produced by Enterobacteriaceae. It induces bacterial filamentation in susceptible cells in a non-SOS-dependent pathway [R. A. Salomon and R. Farias (1992) J. Bacteriol. 174, 7428-7435]. MccJ25 was purified to homogeneity from the growth medium of a microcin-overproducing Escherichia coli strain by reverse-phase HPLC. Based on amino acid composition and absolute configuration determination, liquid secondary ion and electrospray mass spectrometry, extensive two-dimensional NMR, enzymatic and chemical degradations studies, the structure of MccJ25 was elucidated as a 21-residue peptide, cyclo(-Val1-Gly-Ile-Gly-Thr- Pro-Ile-Ser-Phe-Tyr-Gly-Gly-Gly-Ala-Gly-His-Val-Pro-Glu-Tyr-Phe21- ). Although MccJ25 showed high resistance to most of endoproteases, linearization by thermolysin occurred from cleavage at the Phe21-Val1 bond and led to a single peptide, MccJ25-L. While MccJ25 exhibited remarkable antibiotic activity towards Salmonella newport and several E. coli strains (minimal inhibitory concentrations ranging between 0.01 and 0.2 microgram.mL-1), the thermolysin-linearized microcin showed a dramatic decrease of the activity, indicating that the cyclic structure is essential for the MccJ25 biological properties. As MccJ25 is ribosomally synthesized as a larger peptide precursor endowed with an N-terminal extremity, the present study shows that removal of this extension and head-tail cyclization of the resulting propeptide are the only post-translational modifications involved in the maturation of MccJ25, that appears as the first cyclic microcin.  (+info)

Studies on the formation and stability of a complex between Streptomyces proteinaceous metalloprotease inhibitor and thermolysin. (3/552)

The effects of certain physicochemical parameters on the formation and stability of a complex between Streptomyces proteinaceous metalloprotease inhibitor (SMPI) and thermolysin were investigated. SMPI had its lowest Ki value at a pH of around 6.5 (similar to the pH dependence of the kcat/K(m) of thermolysin catalysis), reflecting the splitting mechanism of the SMPI inhibition of thermolysin. This Ki increased with an increase in pressure, and in (Ki-1) was almost linear with respect to pressure. The volume of the reaction (delta Vcomp), which is the volume change accompanying enzyme-inhibitor complex formation, was calculated as +8.1 +/- 0.3 mL.mol-1, which has a sign opposite to delta Vcomp for neutral peptide inhibitors and acyl-peptide substrates. The temperature dependence of Ki-1 gave the reaction enthalpy (delta Hcomp) and reaction entropy (delta Scomp) of the complex formation as 34.6 +/- 1.4 kJ.mol-1 and 298 +/- 5 J.mol-1.K-1, respectively. These positive reaction volumes and reaction entropies were related to the electrostatic interactions and ionic strength dependence of Ki which corresponded to the key ionic interaction during complex formation. Complex formation with SMPI stabilized thermolysin against pressure perturbation as observed by the changes in the Trp fluorescence of thermolysin with increasing pressure. Thermal stability, however, was affected very little by complex formation with SMPI. Phosphoramidon, Cbz-Phe-Gly-NH2 and Cbz-Phe also positively affected the pressure-tolerance of thermolysin, in the following order: Cbz-Gly-Phe-NH2 < Cbz-Phe << phosphoramidon. The third compound exhibited stabilizing effects comparable with those of SMPI, which suggests that the interaction between SMPI and thermolysin was localized to the reactive site.  (+info)

A Kazal-type trypsin inhibitor from the protochordate Ciona intestinalis. (4/552)

A trypsin inhibitor from Ciona intestinalis, present throughout the animal, was purified by ion-exchange chromatography followed by four HPLC steps. By MS the molecular mass of the native form was determined to be 6675 Da. The N-terminal amino acid sequence was determined by protein sequencing, but appeared to be partial because the theoretical molecular mass of the protein was 1101 Da too low. Thermolysin treatment gave rise to several fragments each containing a single disulphide bridge. By sequence analysis and MS intramolecular disulphide bridges could unequivocally be assigned to connect the pairs Cys4-Cys37, Cys8-Cys30 and Cys16-Cys51. The structure of the inhibitor is homologous to Kazal-type trypsin inhibitors. The inhibitor constant, KI, for trypsin inhibition was 0.05 nM whereas chymotrypsin and elastase were not inhibited. To reveal the complete sequence the cDNA encoding the trypsin inhibitor was isolated. This cDNA of 454 bp predicts a protein of 82 amino acid residues including a 20 amino acid signal peptide. Moreover, the cDNA predicts a C-terminal extension of 11 amino acids compared to the part identified by protein sequencing. The molecular mass calculated for this predicted protein is in accordance with the measured value. This C-terminal sequence is unusual for Kazal-type trypsin inhibitors and has apparently been lost early in evolution. The high degree of conservation around the active site strongly supports the importance of the Kazal-type inhibitors.  (+info)

Parvalbumin from rabbit muscle. Isolation and primary structure. (5/552)

A parvalbumin, with its characteristic low molecular weight (approximately 12000) acidic isoelectric point (approximately 5.5), ultraviolet spectrum (maxm 259 nm) and Ca2+-binding capacity (2 mol/mol protein) has been isolated from rabbit (Oryctolagus cuniculus) muscle. Its primary structure has been determined from a study of its tryptic peptides and of overlapping peptides generated by limited tryptic digestion and by chymotryptic and thermolytic digestions of the protein. The amino acid sequence so obtained is considered in comparison with those known for other parvalbumin and for rabbit troponin C.  (+info)

A three-dimensional construction of the active site (region 507-749) of human neutral endopeptidase (EC.3.4.24.11). (6/552)

A three-dimensional model of the 507-749 region of neutral endopeptidase-24.11 (NEP; E.C.3.4.24.11) was constructed integrating the results of secondary structure predictions and sequence homologies with the bacterial endopeptidase thermolysin. Additional data were extracted from the structure of two other metalloproteases, astacin and stromelysin. The resulting model accounts for the main biological properties of NEP and has been used to describe the environment close to the zinc atom defining the catalytic site. The analysis of several thiol inhibitors, complexed in the model active site, revealed the presence of a large hydrophobic pocket at the S1' subsite level. This is supported by the nature of the constitutive amino acids. The computed energies of bound inhibitors correspond with the relative affinities of the stereoisomers of benzofused macrocycle derivatives of thiorphan. The model could be used to facilitate the design of new NEP inhibitors, as illustrated in the paper.  (+info)

The compact conformation of fibronectin is determined by intramolecular ionic interactions. (7/552)

Fibronectin exists in a compact or extended conformation, depending upon environmental pH and salt concentration. Using recombinant fragments expressed in bacteria and baculovirus, we determined the domains responsible for producing fibronectin's compact conformation. Our velocity and equilibrium sedimentation data show that FN2-14 (a protein containing FN-III domains 2 through 14) forms dimers in low salt. Experiments with smaller fragments indicates that the compact conformation is produced by binding of FN12-14 of one subunit to FN2-3 of the other subunit in the dimer. The binding is weakened at higher salt concentrations, implying an electrostatic interaction. Furthermore, segment FN7-14+A, which contains the alternatively spliced A domain between FN11 and 12, forms dimers, whereas FN7-14 without A does not. Segment FN12-14+A also forms dimers, but the isolated A domain does not. These data imply an association of domain A with FN12-14, and the presence of A may favor an open conformation by competing with FN2-3 for binding to FN12-14.  (+info)

Insertion of atToc34 into the chloroplastic outer membrane is assisted by at least two proteinaceous components in the import system. (8/552)

Toc34 is a member of the outer membrane translocon complex that mediates the initial stage of protein import into chloroplasts. Toc34, like most outer membrane proteins, is synthesized in the cytosol at its mature size without a cleavable transit peptide. The majority of outer membrane proteins do not require thermolysin-sensitive components on the chloroplastic surface or ATP for their insertion into the outer membrane. However, different results have been obtained concerning the factors required for Toc34 insertion into the outer membrane. Using an Arabidopsis homologue of pea Toc34, atToc34, we show that the insertion of atToc34 was greatly reduced by thermolysin pretreatment of chloroplasts as assayed either by protease digestion or by alkaline extraction. The insertion was also dependent on the presence of ATP or GTP. A mutant of atToc34 with the GTP-binding domain deleted still required ATP for optimal insertion, indicating that ATP was used by other protein components in the import system. The ATP-supported insertion was observed even in thermolysin-pretreated chloroplasts, suggesting that the protein component responsible for ATP-stimulated insertion is a different protein from the thermolysin-sensitive component that assists atToc34 insertion.  (+info)

To substrater ble brukt i de eksperimetelle studiene. De er referet til som Bradykinin-lignende substrat og AGLA substrat i oppgaven. De eksperimentelle forsøkene gav Km verdi for Bradykinin lignende substrat for Thermolysin fra Bacillus thermoproteolyticus eubakterie på 17.7 +/- 4.3 μM og Pseudolysin fra Pseudomonas aeruginosa på 7.4 +/- 1.3 μM. For AGLA substrat ble Km for Thermolysin målt til 67.7 +/- 10.5 μM, mens for Pseudolysin ble Km målt til 124.8 +/- 22.9 μM. Seksten forbindelser fra en tidligere virtual screening studie av Maybridge databasen viste ingen hemmende effekt hverken på verken Thermolysin eller Pseudolysin. Både under preinkubering av hemmere og under alle forsøk var pH 7.3, mens temperaturen var 37 °C. Av totalt 50 forbindelser (42+8) fra samarbeidspartnere i Italia, viste forbindelse FF33 en IC50 på 754 nM mot Thermolysin og IC50 på 2.28 μM mot Pseudolysin. Forbindelse VDL22 hadde en IC50 på 11,14 μM mot Thermolysin. Forbindelse SM434 viste seg å ...
TY - JOUR. T1 - Kinetic study of thermolysin-catalyzed synthesis of N-(benzyloxycarbonyl)-L-phenylalanyl-L-leucine ethyl ester in an ethyl acetate saturated aqueous system. AU - Nam, K.. AU - Lee, C. K.. AU - Jeong, S. W.. AU - Chi, Y. M.. PY - 2001. Y1 - 2001. N2 - The kinetics of the thermolysin-catalyzed synthesis of N-(benzyloxycarbonyl)-L-phenylalanyl-L-leucine ethyl ester (Z-Phe-LeuOEt) from N-(benzyloxycarbonyl)-L-phenylalanine (Z-Phe) and L-leucine ethyl ester (LeuOEt) in an ethyl acetate saturated aqueous system in a batch operation were studied. The kinetics for the synthesis of Z-Phe-LeuOEt were expressed using a rate equation for the rapid equilibrium random bireactant mechanism. The four kinetic constants involved in the rate equation were determined numerically by the quasi-Newton method so as to fit the calculated results with the experimental data. Within the pH and temperature range examined, the kcat value for the synthesis of Z-Phe-LeuOEt reached a maximum at pH 7.0 and 45°C, ...
Topology analysis of membrane proteins can be obtained by enzymatic shaving in combination with MS identification of peptides. Ideally, such analysis could provide quite detailed information about the membrane spanning regions. Here, we examine the ability of some shaving enzymes to provide large-scale analysis of membrane proteome topologies. To compare different shaving enzymes, we first analyzed the detected peptides from two over-expressed proteins. Second, we analyzed the peptides from non-over-expressed Escherichia coli membrane proteins with known structure to evaluate the shaving methods. Finally, the identified peptides were used to test the accuracy of a number of topology predictors. At the end we suggest that the usage of thermolysin, an enzyme working at the natural pH of the cell for membrane shaving, is superior because: (i) we detect a similar number of peptides and proteins using thermolysin and trypsin; (ii) thermolysin shaving can be run at a natural pH and (iii) the ...
TY - JOUR. T1 - A proteolytic fragment from the central region of p53 has marked sequence-specific DNA-binding activity when generated from wild-type but not from oncogenic mutant p53 protein. AU - Bargonetti, Jill. AU - Manfredi, James J.. AU - Chen, Xinbin. AU - Marshak, Daniel R.. AU - Prives, Carol. PY - 1993. Y1 - 1993. N2 - p53 is a sequence-specific DNA-binding oligomeric protein that can activate transcription from promoters bearing p53-binding sites. Whereas the activation region of p53 has been identified within the amino terminus, the location of the specific DNA-binding domain has not been reported. Thermolysin treatment of p53 protein generates a stable protease-resistant fragment that binds with marked specificity to p53 DNA-binding sites. Amino-terminal sequencing of the fragment located the thennolysin cleavage site to residue 91. Because the fragment does not contain the cdc2 phosphorylation site at Ser-315, we conclude that the the site-specific DNA-binding domain of p53 spans ...
1TRL: NMR solution structure of the C-terminal fragment 255-316 of thermolysin: a dimer formed by subunits having the native structure.
1HYT: Redetermination and refinement of the complex of benzylsuccinic acid with thermolysin and its relation to the complex with carboxypeptidase A.
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Looking for Inouye, Kaoru? Find out information about Inouye, Kaoru. 1835-1915, Japanese statesman. He was a leader of the antiforeign movement in his native Choshu fief, and helped set fire to the British legation in Edo in... Explanation of Inouye, Kaoru
Variants of this enzyme have been found in species of Bacillus including B. subtilis [1,6], B. amyloliquefaciens [5], B. megaterium (megateriopeptidase, [2]), B. mesentericus [10], B. cereus [3,8,9] and B. stearothermophilus [7]. In peptidase family M4 (thermolysin family). Formerly included in EC 3.4.24.4 ...
SWISS-MODEL Template Library (SMTL) entry for 1hyt.1. RE-DETERMINATION AND REFINEMENT OF THE COMPLEX OF BENZYLSUCCINIC ACID WITH THERMOLYSIN AND ITS RELATION TO THE COMPLEX WITH CARBOXYPEPTIDASE A
SWISS-MODEL Template Library (SMTL) entry for 5tac.1. Conformational Sampling Differences across the Arrhenius Plot Biphasic Break Point at Ambient Temperature in the Enzyme Thermolysin
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TY - JOUR. T1 - 3D-QSAR of Angiotensin-Converting Enzyme and Thermolysin Inhibitors. T2 - A Comparison of CoMFA Models Based on Deduced and Experimentally Determined Active Site Geometries. AU - DePriest, Scott A.. AU - Mayer, Dorica. AU - Naylor, Christopher B.. AU - Marshall, Garland R.. AU - Mayer, Dorica. AU - Naylor, Christopher B.. PY - 1993/6/1. Y1 - 1993/6/1. N2 - The ability of comparative molecular field analysis (CoMFA), a three-dimensional, quantitative structure-activity relationship (3-D QSAR) paradigm, to predict the activity of inhibitors of angiotensin-converting enzyme (ACE) and thermolysin was examined. Correlations derived from computationally and experimentally determined alignment rules were compared. The correlations derived for the ACE series using alignment rules determined from a systematic conformational search (Mayer, D.; Naylor, C. B.; Motoc, I.; Marshall, G. R. J. Comput.-Aided Molec. Des. 1987, 1, 3-16) were comparable to those derived for the thermolysin ...
Ungaro, V. A., Fairbanks, J. P. A., Rossi, L. M., & Machini, M. T. (2017). Thermolysin immobilized on Fe IND. 3O IND. [email protected] nanoparticle: preparation and characterization of a new recoverable biocatalyst. In Proceedings. Durham: International Union of Pure and Applied Chemistry (IUPAC). Recuperado de http://www.neopixdmi.com.br/@mci/iupac2017 ...
Hereditary mutations in the transforming growth factor beta induced (TGFBI) gene cause phenotypically distinct corneal dystrophies characterized by protein deposition in cornea. We show here that the Arg555Trp mutant of the fourth fasciclin 1 (FAS1-4) domain of the protein (TGFBIp/keratoepithelin/betaig-h3), associated with granular corneal dystrophy type 1, is significantly less susceptible to proteolysis by thermolysin and trypsin than the WT domain. High-resolution liquid-state NMR of the WT and Arg555Trp mutant FAS1-4 domains revealed very similar structures except for the region around position 555. The Arg555Trp substitution causes Trp555 to be buried in an otherwise empty hydrophobic cavity of the FAS1-4 domain. The first thermolysin cleavage in the core of the FAS1-4 domain occurs on the N-terminal side of Leu558 adjacent to the Arg555 mutation. MD simulations indicated that the C-terminal end of helix alpha3 containing this cleavage site is less flexible in the mutant domain, ...
Enzymatic hydrolysis of proteins is used to improve nutritional and functional properties of many foods. The desired degree of hydrolysis (DH) depends on food application. Effective hydrolysis requires optimal hydrolysis conditions for both the enzyme and the substrate protein. This study aimed to hydrolyze the oat globulins (OG) effectively under different conditions. Our first goal was to maximise the OG solubility and then to hydrolyze OG under optimised conditions. The solubility of isolated OG in Na-phosphate solutions containing 0 1 M NaCl was determined. Globulins were subjected to single-enzyme hydrolysis with either subtilisin, thermolysin or pepsin. In addition, OG were degraded in two-stage hydrolysis first with pepsin and then either with subtilisin or thermolysin. The hydrolysates were analysed by SDS-PAGE and DH was quantified with the OPA method. The solubility of OG increased when NaCl was added at pH 5 10. Under more acidic conditions the solubility, however, decreased with ...
0047] The above characteristics are optionally taken into account when producing a protein with reduced or improved enzymatic activity. Illustratively, substitutions in a substrate binding site, exosite, cofactor binding site, catalytic site, or other site in an enzyme may alter the activity of the enzyme toward a substrate. In considering such substitutions the sequences of other known naturally occurring or non-naturally occurring proteins may be taken into account. Illustratively, mutations of L134R and S320A in Bacillis licheniformis α-amylase improve the catalytic activity of the enzyme in acidic conditions 14-fold. Liu, et al., Appl Microbiol Biotechnol, 2008; 80:795-803. As another example, a corresponding mutation to that of Asp213 in thermolysin is operable such as that done by Mild, Y, et al., Journal of Molecular Catalysis B: Enzymatic, 1996; 1:191-199. Optionally, a substitution in thermolysin of L144S alone or along with substitutions of G8C/N60C/S65P are operable to increase the ...
Aceetate [ ACEETATE, n. [See Acid.] In chimistry, a neutral salt formed by the union of the acetic acid, or radical vinegar, with any salifiable base, as with earths, metals, and alkalies; as the acetate of alumine, of lime, or of copper. ]
Is able to inhibit all four classes of proteinases by a unique trapping mechanism. This protein has a peptide stretch, called the bait region which contains specific cleavage sites for different proteinases. When a proteinase cleaves the bait region, a conformational change is induced in the protein which traps the proteinase. The entrapped enzyme remains active against low molecular weight substrates (activity against high molecular weight substrates is greatly reduced). Following cleavage in the bait region a thioester bond is hydrolyzed and mediates the covalent binding of the protein to the proteinase (By similarity). Displays inhibitory activity against chymotrypsin, papain, thermolysin, subtilisin A and, to a lesser extent, elastase but not trypsin. May play an important role during desquamation by inhibiting extracellular proteases ...
Lysates from logarithmic growth phase T. brucei (2.5 × 108 cells) isolated from infected rats and from the logarithmic growth and late stationary phase (8.3 and 5.4 × 107 cells, respectively) organisms isolated from bloodstream-form cultures were prepared by hypotonic lysis using double-distilled water containing a cocktail of reversible and irreversible inhibitors (one tablet in 25 ml) of pancreas extract, pronase, thermolysin, chemotrypsin, trypsin, and papain (Complete™; Roche Diagnostics). For PG production from AA, we used the reaction mixture described by Ujihara et al. (21) with the following modifications: 100 mM sodium phosphate, pH 7.0, 2 μM hematin, 5 mM tryptophan, 1 mM AA, and 300 μl of the respective T. brucei lysates in a final volume of 500 μl. The mixture was incubated at 37°C for 30 min, and then the reaction was stopped by addition of 100 μl of 1 M HCl and 6 vol of cold ethyl acetate.. For PGF2α synthesis from PGH2, a standard reaction mixture that contained 100 mM ...
Methyl, Ethyl, Propyl, Butyl: Futile But Not for Water, as the Correlation of Structure and Thermodynamic Signature Shows in a Congeneric Series of Thermolysin Inhibitors ...
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Enzymatische Spaltung nativer cystinhaltiger Polypeptide durch Thermolysin (E.S. 3.4.4.), II: Vergleich von Thermolysin mit α-Protease aus Crotalus atrox-Gift und Subtilisin (Enzymic hydrolysis of native cystine-containing polypeptides with thermolysin. II. Comparison of thermolysin with α-protease from Crotalus atrox poison and subtilisin) ...
Thermostable enzyme from Vibrio proteolyticus (formerly Aeromonas proteolytica). Specificity related to, but distinct from, those of thermolysin and bacillolysin [1]. A zinc metallopeptidase in family M4 (thermolysin family). Formerly included in EC 3.4.24.4 ...
2-{[3-(3,4-Dihydroxyphenyl)acryloyl]amino}benzoic acid | C16H13NO5 | CID 54039925 - structure, chemical names, physical and chemical properties, classification, patents, literature, biological activities, safety/hazards/toxicity information, supplier lists, and more.
Rabbit monoclonal antibody raised against a human MKI67IP peptide using ARM Technology. A synthetic peptide of human MKI67IP is used for rabbit immunization.Customer or Abnova will decide on the preferred peptide sequence. (H00084365-K) - Products - Abnova
Over 70 metallopeptidase families have been identified to date. In these enzymes a divalent cation which is usually zinc, but may be cobalt, manganese or copper, activates the water molecule. The metal ion is held in place by amino acid ligands, usually three in number. In some families of co-catalytic metallopeptidases, two metal ions are observed in crystal structures ligated by five amino acids, with one amino acid ligating both metal ions. The known metal ligands are His, Glu, Asp or Lys. At least one other residue is required for catalysis, which may play an electrophillic role. Many metalloproteases contain an HEXXH motif, which has been shown in crystallographic studies to form part of the metal-binding site [(PUBMED:7674922)]. The HEXXH motif is relatively common, but can be more stringently defined for metalloproteases as abXHEbbHbc, where a is most often valine or threonine and forms part of the S1 subsite in thermolysin and neprilysin, b is an uncharged residue, and c a ...
Inouye et al used some simple questions to screen patients for their level of social support. The authors are from Yale University in New Haven, Connecticut.
In 1 we will find the explanation of clinical results obtained by intravenous CH1 injections, even if the blood pH did not change.. In 2 we find the reason for clinical results obtained by increasing neutral salts intake. Ambard gives the following reaction:. NaCl+2CO3 NaHCO3+HCl. Whereas this reaction gives birth to infinitesimal quantities of HCl, if it takes place in the presence of albumin, the acid will impregnate the albumin and a new quantity of NaCl can be decomposed and liberates a new fraction of chlorine.. Here then we have first of all a new characteristic of proteins. It is therefore possible to perceive, that though the proteins are charged with acid, the medium many manifest a neutral reaction. Chabanier and Lobo-Onell have shown that even an increase of acid, producing an acidemia (of the medium), will allow the albumins to discharge themselves of HCl if the neutral salt content is diminished. It is possible, therefore, to have an acidemia (of the medium) and an alkalosis (of the ...
In well-known methods of estimating rates of irreversible disposal (utilization) in vivo the rates are calculated from the areas to infinity under specific radioactivity-time (S-t) or quantity-of-label-time (q-t) curves obtained by measurements on samples of plasma after intravenous injection of labelled substrate. The errors in the calculated rates are mostly those of the estimates of the areas. These errors are of two kinds: random, caused by the variances of the values of S or q, and systematic, caused by differences between the curves used to interpolate between these values and the true curves. A rigorous method is given for calculating the random errors from the variances of the values of S or q, and is applied to choosing the best times to sample the plasma from small animals from which few plasma samples can be taken. A procedure for estimating systematic errors is also given. Programs in BASIC language to carry out the calculations are deposited as Supplementary Publication SUP 50058 (5 ...
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It has been designed as a DIRECT REPLACEMENT in units such as the ZR33, Magnum Club Smoke, AF1 MkI, Magnum 2000 DMX interface, ZR12 DMX interface etc ...
The Liberase projects showed that collagenase and neutral protease were the enzymes required for the release of islets from human pancreatic tissue. Liberase contains purified class I and class II C histolyticum collagenase, and thermolysin or Dispase. Different formulations and doses were used to isolate cells from different tissues.. The identification of the C. histolyticum genes and corresponding protein domain structures of class I and class II collagenase provided new insights into the mechanism of enzyme-mediated tissue dissociation. Both classes of collagenase contain four protein domains: a large catalytic domain that cuts native collagen or gelatin, linking domain(s) (no known function), and collagen binding domain(s). Intact class I has one catalytic domain, a linking domain, and two collagen binding domains whereas intact class II has a catalytic domain, two linking domains, and one collagen binding domain. Only those forms of collagenase containing a catalytic domain and at least ...
The Liberase projects showed that collagenase and neutral protease were the enzymes required for the release of islets from human pancreatic tissue. Liberase contains purified class I and class II C histolyticum collagenase, and thermolysin or Dispase. Different formulations and doses were used to isolate cells from different tissues.. The identification of the C. histolyticum genes and corresponding protein domain structures of class I and class II collagenase provided new insights into the mechanism of enzyme-mediated tissue dissociation. Both classes of collagenase contain four protein domains: a large catalytic domain that cuts native collagen or gelatin, linking domain(s) (no known function), and collagen binding domain(s). Intact class I has one catalytic domain, a linking domain, and two collagen binding domains whereas intact class II has a catalytic domain, two linking domains, and one collagen binding domain. Only those forms of collagenase containing a catalytic domain and at least ...
Here we present the synthesis and post-polymerisation modification of poly(acryloyl hydrazide), a versatile scaffold for the preparation of functional polymers: poly(acryloyl hydrazide) was prepared from commercially available starting materials in a three step synthesis on a large scale, in good yields and high purity. Our synthetic approach included the synthesis of a Boc-protected acryloyl hydrazide, the preparation of polymers via RAFT polymerisation and the deprotection of the corresponding Boc-protected poly(acryloyl hydrazide). Post-polymerisation modification of poly(acryloyl hydrazide) was then demonstrated using a range of conditions for both hydrophilic and hydrophobic aldehydes. These experiments demonstrate the potential of poly(acryloyl hydrazide) as a scaffold in the synthesis of functional polymers, in particular those applications where in situ screening of the activity of the functionalised polymers may be required (e.g. biological applications ...
Page contains details about poly(D-α-tocopheryl polyethylene glycol 1000-2,2-bis(acryloyloxymethyl)propionate-co-N,N-bis(acryloyl)cystamine-co-1,6-hexanediol diacrylate-co-piperazine) nanoparticles in phosphate-buffered saline (pH 6.5) . It has composition images, properties, Characterization methods, synthesis, applications and reference articles : nano.nature.com
0022]The (meth)acryloyl groups in the linear (meth)acryloyl-containing compound of the present invention exist at parts of side chains relative to the molecular chain having the longest coupling length in a single molecule, and it is indispensable from the standpoint of satisfactory hardenability and sensitivity that an average of 3 or more be contained in a single molecule. With respect to the pertinent (meth)acryloyl groups, by selecting the structural units which configure the straight-chain portion, it is possible to easily adjust their concentration in a single molecule, that is, the molecular weight per (meth)acryloyl group. For example, when using residue excluding 2 hydroxyl groups from bisphenol as A in the aforementioned general formula (1) as described below, it is possible to obtain approximately the same concentration as with conventional bifunctional epoxy(meth)acrylate, and to have crosslink density in an appropriate range while having multifunctionality, thereby obtaining ...
Note: For nodular (composite) ganglioneuroblastomas with more than 1 nodule, degree of differentiation and mitotic-karyorrhectic index (MKI) must be given for each nodule. Please indicate the differentiation and MKI for the least favorable nodule in the checklist below. Classification of additional nodules can be described in the Comment. ...
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All text is © British Library Board and is available under a Creative Commons Attribution Licence, except where otherwise stated.. ...
All text is © British Library Board and is available under a Creative Commons Attribution Licence, except where otherwise stated.. ...
All text is © British Library Board and is available under a Creative Commons Attribution Licence, except where otherwise stated.. ...
A SU-30 MKI combat jet belonging to the IAF crashed in the Pokhran firing range in Rajasthan during a firepower demonstration drill.
Take part in our web survey!. Why not take a few moments to tell us what you think of our website?. Your views could help shape our site for the future.. Choose Yes please to open the survey in a new browser window or tab, and then complete it when you are ready. ...
This is a chart of the English Channel possibly dating from 1542-1544. It shows the south coast of England and the facin.... ...
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Kresge, Nicole; Simoni, Robert D.; Hill, Robert L. (2009). "Structural Studies of Thermolysin: the Work of Brian W. Matthews". ... Matthews BW, Colman PM, Jansonius JN, Titani K, Walsh KA, Neurath H (1972). "Structure of thermolysin". Nature New Biology. 238 ... He also solved early structures of the thermophilic bacterial enzyme thermolysin, the helix-turn-helix DNA-binding ...
The catalytic site is contained within a thermolysin-like region found in many metallopeptidases and located in the domain near ... Matthews BW, Weaver LH, Kester WR (December 1974). "The conformation of thermolysin". The Journal of Biological Chemistry. 249 ...
It is an inhibitor of the enzyme thermolysin, a membrane metallo-endopeptidase inhibitor, and an endothelin converting enzyme ... "Binding between thermolysin and its specific inhibitor, phosphoramidon". Journal of Biochemistry. 95 (2): 529-34. PMID 6715312 ...
The protease thermolysin can be fully inactivated by EDTA. This feature of thermolysin makes FASTpp compatible with subsequent ... in a thermal gradient PCR cycler to different maximal temperatures in presence of the thermostable protease thermolysin (see ...
"Circular dichroism comparative studies of two bacterial collagenases and thermolysin". Biochimica et Biophysica Acta (BBA) - ...
It is a known reversible inhibitor of thermolysin and is expected to inhibit other metalloproteinases. Chemically, talopeptin ... Kitagishi, Keiko; Hiromi, Keitaro (1984). "Inhibitory spectrum of talopeptin (MKI), a specific inhibitor of thermolysin". ... "Equilibrium study on the binding between thermolysin and Streptomyces metalloprotease inhibitor, talopeptin (MKI)". Journal of ...
Alternatively, it has been shown for thermolysin (another metalloproteinase) that the amide product can be released in its ... Hangauer DG, Monzingo AF, Matthews BW (Nov 1984). "An interactive computer graphics study of thermolysin-catalyzed peptide ... "A theoretical study of the mechanism for peptide hydrolysis by thermolysin". Journal of Biological Inorganic Chemistry. 7 (3): ...
Thermolysin complexed with the inhibitor (S)-thiorphan are isomeric thiol-containing inhibitors of endopeptidase EC 24-11 (also ... The peptidase domain for members of this family also contains a bacterial member and resembles that of thermolysin the ... "Thiorphan and retro-thiorphan display equivalent interactions when bound to crystalline thermolysin". Biochemistry. 28 (4): ... predicted active site residues for members of this family and thermolysin occur in the motif HEXXH. ...
"Extreme stabilization of a thermolysin-like protease by an engineered disulfide bond". The Journal of Biological Chemistry. 272 ...
2006). "A quenched fluorescent dipeptide for assaying dispase- and thermolysin-like proteases". Analytical Biochemistry. 352 (1 ...
Some silanediols and silanetriols inhibit hydrolytic enzymes such as thermolysin and acetycholinesterase. Literally, silanol ...
Thermolysin - cuts before Ile, Met, Phe, Trp, Tyr, or Val, unless preceded by Pro. Sometimes cuts after Ala, Asp, His or Thr. ...
The overall topology of this domain is shared by the archetypal zinc-endopeptidase thermolysin. Astacin protease domains also ... the type example for clan MA and the predicted active site residues for members of this family and thermolysin occur in the ... The protein fold of the peptidase domain for members of this family resembles that of thermolysin, ... molecular structure and comparison with thermolysin". Journal of Molecular Biology. 229 (4): 945-68. doi:10.1006/jmbi.1993.1098 ...
"Substrate recognition and selectivity of peptide deformylase Similarities and differences with metzincins and thermolysin". J. ...
Grobelny D, Poncz L, Galardy RE (August 1992). "Inhibition of human skin fibroblast collagenase, thermolysin, and Pseudomonas ... Examples of enzymes that ilomastat inhibit include rabbit MMP9, thermolysin, peptide deformylase, and anthrax lethal factor ...
Lipases, thermolysin, galactosidase, nucleases, and trypsin have all been used in the removal of cells. After a cell is lysed ...
Kester WR, Matthews BW (1977). "Crystallographic study of the binding of dipeptide inhibitors to thermolysin: implications for ...
This enzyme catalyses the following chemical reaction Similar, but not identical, to that of thermolysin This enzyme is present ... "The primary structure of Bacillus cereus neutral proteinase and comparison with thermolysin and Bacillus subtilis neutral ... from Bacillus cereus refined at 3.0 A resolution and comparison with the homologous but more thermostable enzyme thermolysin". ...
Joudiou C, Carvalho KM, Camarao G, Boussetta H, Cohen P (June 1993). "Characterization of the thermolysin-like cleavage of ...
Thermolysin is an enzyme produced by Bacillus thermoproteolyticus that catalyses the hydrolysis of peptides containing ... Holden, Hazel; Tronrud, D. E.; Monzingo, A. F.; Weaver, L. H. (1987). "Slow-and fast-binding inhibitors of thermolysin display ... mechanism starts form the small peptide molecule and replaces the zinc binding water molecule towards Glu143 of thermolysin. ...
However, commercially available thermolysin is dependent on calcium ions for activity and denatures itself just above 85 ... In fast parallel proteolysis the researcher adds a thermostable protease (thermolysin) and takes out samples in parallel upon ... FastPP exploits that proteins become increasingly susceptible to proteolysis when unfolded and that thermolysin cleaves at ... showed that stabilization by ligand binding could impart resistance to proteolytic digestion with thermolysin. Protection ...
... may refer to: Subtilisin, an enzyme Thermolysin, an enzyme This disambiguation page lists articles associated with ...
Holland, D.R.; Barclay, P.L.; Danilewicz, J.C.; Matthews, B.W.; James, K. (January 1994). "Inhibition of thermolysin and ... the gluzincins a faint but significant structural relationship of the metzincins to the thermolysin-like enzymes, Zincin is the ... neutral endopeptidase 24.11 by a novel glutaramide derivative: X-ray structure determination of the thermolysin-inhibitor ...
... which distinguished this enzyme from thermolysin This thermostable enzyme is isolated from Vibrio proteolyticus. Holmquist B, ...
... involvement of a thermolysin-like metalloendopeptidase (enkephalinase), angiotensin-converting enzyme, and other unidentified ...
It has 102 amino acid residues with two disulphide bridges and specifically inhibits metalloproteinases such as thermolysin, ...
It was initially produced as a mixture made by thermolysin digestion of protamine, but the actual effective peptide portion ...
"Isolation and amino acid sequence of a peptide containing an epoxide-reactive residue from the thermolysin-digest of ...
... thermolysin family). Morihara K, Tsuzuki H (1975). "Pseudomonas aeruginosa elastase: affinity chromatography and some ... Insulin B chain cleavage pattern identical to that of thermolysin, but specificity differs in other respects This enzyme ...
... its active site is homologous to those of well studied zinc-proteases such as carboxypeptidase A and thermolysin. Therefore, it ...
However thermolysin is also widely used for peptide bond formation through the reverse reaction of hydrolysis. Thermolysin is ... Like all bacterial extracellular proteases thermolysin is first synthesised by the bacterium as a pre-proenzyme. Thermolysin is ... Thermolysin has a T50 value of 86.9 °C, making it the most thermo stable member of the TLP family. Studies on the contribution ... Thermolysin has a molecular weight of 34,600 Da. Its overall structure consists of two roughly spherical domains with a deep ...
This lyophilized enzyme is from Bacillus thermoproteolyticus. This enzyme is characterized by its excellent heat stability and substrate specificity towards isoleucine, methionine and valine. Activity: 7000 PU/mg of protein.
Thermolysin 1427 3.4.24.27 , Details 100 2WHZ 1 A THERMOLYSIN THE DI-PEPTIDE IY IS BOUND TO THE PROTEIN 1427 3.4.24.27 , ... Thermolysin 1427 3.4.24.27 , Details 138 2WI0 1 A THERMOLYSIN THE DI-PEPTIDE LW IS BOUND TO THE PROTEIN 1427 3.4.24.27 , ... Thermolysin UNP residues 233-548 1427 3.4.24.27 , Details 27 5M5F 1 E Thermolysin UNP residues 233-548 1427 3.4.24.27 , Details ... Thermolysin UNP residues 233-548 1427 3.4.24.27 , Details 15 4MWP 1 E Thermolysin Mature form (UNP residues 233-548) 1427 3.4. ...
Conformational Constraint and Structural Complementarity in Thermolysin Inhibitors: Structures of Enzyme Complexes and ...
Similarities and differences with metzincins and thermolysin.. Ragusa S1, Mouchet P, Lazennec C, Dive V, Meinnel T. ... Based on the numerous homologies that deformylase displays with thermolysin and metzincins, a mechanism of enzyme:substrate ...
Model building of a thermolysin-like protease by mutagenesis. Frigerio, F., Margarit, N. V., Nogarotto, R., Grandi, G., Vriend ... thermolysin) and Bacillus cereus as templates, The largest portion of NP-sub could be modelled satisfactorily, using standard ...
In this article, we use four alternative proteases: Arg-C, elastase, thermolysin and pepsin, and show typical applications for ... The Elastase, Thermolysin and Pepsin Advantage. Elastase (Cat.# V1891), thermolysin (Cat.# V4001) and pepsin (Cat.# V1959) are ... Thermolysin and pepsin. Thermolysin and pepsin are distinct from other proteases because they tolerate extreme conditions: high ... Protein digestion with thermolysin is rapid because the reaction occurs at a higher temperature. However, when high temperature ...
iv) Fewer detected peptides from thermolysin shaving originate from the transmembrane regions. Using thermolysin shaving we can ... we detect a similar number of peptides and proteins using thermolysin and trypsin; (ii) thermolysin shaving can be run at a ... At the end we suggest that the usage of thermolysin, an enzyme working at the natural pH of the cell for membrane shaving, is ... Membrane protein shaving with thermolysin can be used to evaluate topology predictors. Bendz, Maria ...
... based on specific tagging with biotin or proteolysis using thermolysin, a non-membrane permeable protease. To evaluate this ... based on specific tagging with biotin or proteolysis using thermolysin, a non-membrane permeable protease. To evaluate this ... away by the thermolysin treatment, whereas IEP37 and MGD1 were still detected in thermolysin-treated samples (Figures 4B-D, ... At this stage, thermolysin treatment or biotin tagging was performed as described below. Envelope, thylakoids, and stroma were ...
Inhibition of Bacillus thermoproteolyticus thermolysin using FA-Fly-Leu-NH2 as substrate at pH 7.5 after 15 mins by Henderson ...
However, activity of thermolysin was abolished in all organic solvents. In the presence of DTT 1mM, the PAE activity was ... Thermolysin from Bacillus thermoproteolyticus is a highly thermostable Zn-metalloprotease and is applied in industry for ... However, thermolysin has low stability at organic solvents.In contrast, its homologous enzyme, Pseudomonas aeruginosa elastase ... In the present study, recombinant PAE was purified and compared with thermolysin in different organic solvents. For this ...
Fitzpatrick, Paul A. (2007) Shear stress-mediated regulation of thermolysin-like zinc metallopeptidase expression levels in ... Shear stress-mediated regulation of thermolysin-like zinc metallopeptidase expression levels in vascular endothelial cells ...
... thermolysin (EC 3.4.24.27). One active site residue of thermolysin, Arg-203, is involved in inhibitor binding by forming ... In addition, the replacement of Asp-170 of thermolysin by Ala residue resulted in a decrease in kcat/Km of 220-fold. The ... This led, in both cases, to decreases in kcat/Km values, of 122-fold for neprilysin and 2300-fold for thermolysin, essentially ... Sequence alignment data shows that Arg-717 of neprilysin could play a similar role to Arg-203 of thermolysin. This was ...
Thermolysin \ 3504 for more molecular products just contact us ... Thermolysin (EC 3.4.24.27). [SCATT_p12040] Thermolysin. [nprS ... OV320_0165] Thermolysin (EC 3.4.24.27). [OV320_1504] Thermolysin (EC 3.4.24.27). [OV450_3101] Thermolysin (EC 3.4.24.27). [npr2 ... npr2 BT246_27540] Thermolysin (EC 3.4.24.27). [npr2 BTGOE6_18890] Thermolysin (EC 3.4.24.27). [npr2_1 BTGOE7_18030] Thermolysin ... npr2 BTI247_27420] Thermolysin (EC 3.4.24.27). [nprS SsS58_07371] Thermolysin (EC 3.4.24.27). [MME] Neprilysin (EC 3.4.24.11) ( ...
THERMOLYSIN (70% ACETONE SOAKED CRYSTALS). Components. THERMOLYSIN. Details. Keywords. HYDROLASE / METALLOPROTEINASE / ORGANIC ... THERMOLYSIN. / Details. Sequence. Seq. region. Function. Str. site (34). Validation (3). Mass: 34362.305 Da / Num. of mol.: 1 ... thermolysin / metalloendopeptidase activity / extracellular region / metal ion binding. Peptidase M4/M1, CTD superfamily / ... Details: 4.6MM THERMOLYSIN, 45% DMSO, 50MM TRIS-HCL, 2.5M CSCL, pH 7.5, VAPOR DIFFUSION, SITTING DROP, temperature 298K. ...
title = "Reversible acetonitrile-induced inactivation/activation of thermolysin",. abstract = "Thermolysin is catalytically ... Reversible acetonitrile-induced inactivation/activation of thermolysin. R.V. Ulijn, A.E.M. Janssen, B.D. Moore, P.J. Halling, S ... Reversible acetonitrile-induced inactivation/activation of thermolysin. / Ulijn, R.V.; Janssen, A.E.M.; Moore, B.D.; Halling, P ... N2 - Thermolysin is catalytically inactive in mixtures or 70-15% acetonitrile in aqueous buffer. Unexpectedly, dilution of the ...
Colman PM, Jansonius JN, Matthews BW (1972) The structure of thermolysin: an electron density map at 2-3 Å resolution. J Mol ... Thermolysins constitute a family of secreted bacterial metalloproteases expressed, among others, by several pathogens. Strains ... The internal region contains the HEXXH catalytic domain that is conserved in members of the thermolysin family. Regardless of ... Buchanan JD, Corbett RJ, Roche RS (1986) The thermodynamics of calcium binding to thermolysin. Biophys Chem 23: 183-199. ...
Thermolysin complexed with Z-L-Glutamic acid (benzyloxycarbonyl-L-Glutamic acid) ... Thermolysin complexed with Z-L-Glutamic acid (benzyloxycarbonyl-L-Glutamic acid). Coordinates. PDB Format Method. X-RAY ... Senda, M. et al., Crystal structure analyses of thermolysin in complex with its inhibitors. To be Published Release Date. 2002- ...
Experimental MAD phased structure of thermolysin in complex with inhibitor JC65. ... Experimental MAD phased structure of thermolysin in complex with inhibitor JC65.. Coordinates. PDB Format Method. X-RAY ...
Thermolysin This lyophilized enzyme is from Bacillus thermoproteolyticus. This enzyme is characterized by its excellent heat ...
Cut sites for Elastase, Pepsin and Thermolysin.. Cut sites for Elastase, Pepsin and Thermolysin.. ... Thermolysin. Cleavage sites: N-terminal of Leu, Phe, Val, Ala, Met, Ile. Optimal pH: 5-8.5. ...
4 Additional thermolysin digestions of brain homogenates from inoculated TgM83 mice.. a) Immunoblots of detergent-insoluble α- ... a) Thermolysin digestion of brain extracts from the three human synucleinopathy samples inoculated into TgM83 mice. α-Syn was ... with or without digestion with thermolysin (TL). Brain homogenates from asymptomatic TgM83 mice from the second passage of PBS ...
Thermolysin Bacillus thermoproteolyticus 0.992 CHEMBL236 Delta opioid receptor Homo sapiens 0.991 CHEMBL5857 Trace amine- ...
25 μg of thermolysin per mg of protein (+ thermolysin), (iii) 10 mM EDTA and 25 μg of thermolysin per mg of protein (inhibited ... thermolysin), and (iv) 0.1% Triton X-100 and 25 μg of thermolysin per mg of protein (permeabilized organelles + thermolysin). ... Intact apicoplasts were incubated with thermolysin, then examined by Western blotting (28). Thermolysin treatment of intact ... D) Thermolysin digestion of free, intact apicoplasts from parasites transfected with an N-terminally tagged copy of PfoTPT (HA- ...
Owusu R, Doble C. Heat-stability of a proteinase from psychrotrophic Pseudomonas fluorescens P38, chymotrypsin and thermolysin ... Heat-stability of a proteinase from psychrotrophic Pseudomonas fluorescens P38, chymotrypsin and thermolysin. Richard Owusu, C ... Heat-stability of a proteinase from psychrotrophic Pseudomonas fluorescens P38, chymotrypsin and thermolysin. / Owusu, Richard ... title = "Heat-stability of a proteinase from psychrotrophic Pseudomonas fluorescens P38, chymotrypsin and thermolysin", ...
Enzymatische Spaltung nativer cystinhaltiger Polypeptide durch Thermolysin (E.S. 3.4.4.) / Enzymatic Hydrolysis of Native ...
... thermolysin; and nuclease (RNAase and DNAase).Partially degraded by pepsin and papain.______________________________________. ...
Thermolysin. Protease from Bacillus thermoproteolyticus. 14 U/mg. pH 7; 50 °C. ... The BLG was hydrolysed by neutrase (Neu), alcalase (Alc), savinase (Sav), elastase (Ela), thermolysin (Ther), and trypsin (Try ... Thermolysin (Ther), and Savinase (Sav). The statistical analysis of the results was performed by two-way ANOVA followed by a ... Thermolysin (Ther), and Savinase (Sav). The statistical analysis of the results was performed by two-way ANOVA followed by a ...
Thermolysin in human cultured keratinocyte isolation Gragnani, A; Sobral, C. S; Ferreira, L. M. *Gragnani, A; Federal ... Full text: Available Index: LILACS (Americas) Main subject: Thermolysin / Trypsin / Keratinocytes / Cell Separation / Colony- ... Full text: Available Index: LILACS (Americas) Main subject: Thermolysin / Trypsin / Keratinocytes / Cell Separation / Colony- ... To compare the action of trypsin and thermolysin in the keratinocyte isolation using newborn foreskin. METHODS:. This method ...
... and the metallopeptidase thermolysin. The substrates were successfully used to monitor the activities of cysteine peptidases ...
  • Thermolysin (EC 3.4.24.27, Bacillus thermoproteolyticus neutral proteinase, thermoase, thermoase Y10, TLN) is a thermostable neutral metalloproteinase enzyme produced by the Gram-positive bacteria Bacillus thermoproteolyticus. (wikipedia.org)
  • Thermolysin is the most stable member of a family of metalloproteinases produced by various Bacillus species. (wikipedia.org)
  • Changing these amino acids to threonine (T) and alanine (A) respectively in a less stable thermolysin-like proteinase produced by Bacillus stearothermophillus (TLP-ste), results in individual reductions in stability of 7 °C (F63→T) and 6.3 °C (P69→A) and when combined a reduction in stability of 12.3 °C. In the synthesis of aspartame, less bitter-tasting byproduct is produced when the reaction is catalyzed by thermolysin. (wikipedia.org)
  • 1976). "Role of calcium ions in the thermostability of thermolysin and Bacillus subtilis var. (wikipedia.org)
  • Thermolysin from Bacillus thermoproteolyticus is a highly thermostable Zn-metalloprotease and is applied in industry for peptide synthesis, a reaction essentially performs in organic media. (magiran.com)
  • De eksperimentelle forsøkene gav Km verdi for Bradykinin lignende substrat for Thermolysin fra Bacillus thermoproteolyticus eubakterie på 17.7 +/- 4.3 μM og Pseudolysin fra Pseudomonas aeruginosa på 7.4 +/- 1.3 μM. (uit.no)
  • We sought a method providing information at the surface of the outer envelope membrane (OEM), based on specific tagging with biotin or proteolysis using thermolysin, a non-membrane permeable protease. (frontiersin.org)
  • However, thermolysin has low stability at organic solvents.In contrast, its homologous enzyme, Pseudomonas aeruginosa elastase (PAE) is an organic solvent stable protease. (magiran.com)
  • 8 of these 5 mL samples were prepared, and each of the four proteases (proteinase K, trypsin, chymotrypsin, and thermolysin) were added in the amounts specified in their respective reaction protocols to achieve final protease concentrations of 1 and 100 nM. (openwetware.org)
  • The serine protease ALP-901 produced the highest yield after 48 hours reaction time (53 %), while the reaction with thermolysin achieved the overall highest yield (63 %) after 6 hours, with only monoesters synthesised. (aau.dk)
  • The present invention provides methods and compositions comprising at least one thermolysin-like neutral protease enzyme with improved storage stability and/or. (patents.com)
  • HA/protease belongs to the thermolysin family of bacterial zinc metalloproteases ( 36 ). (asm.org)
  • Like all bacterial extracellular proteases thermolysin is first synthesised by the bacterium as a pre-proenzyme. (wikipedia.org)
  • Based on the assumption that leptospiral proteases play a crucial role during infection, the aim of this work was to obtain a functional recombinant thermolysin for future studies on the role of these metalloproteases on leptospiral infection. (butantan.gov.br)
  • Beynon, R.J. & Beaumont, A. (1998) M4 proteases: thermolysins. (liverpool.ac.uk)
  • The enzymes reviewed include collagenase, trypsin, clostripain, and the neutral proteases dispase and thermolysin. (mendeley.com)
  • The heat-stabilities of proteinases from a psychrotrophic Psuedomonas fluorescens strain P38 (P38 proteinase), chymotypsin, succinyl-chymotrypsin and thermolysin are compared on the basis of their respective thermoinactivation-rate constants and Arrhenius plots (40-145°C). The Arrhenius plots for P38 proteinase and succinyl-chymotrypsin showed an inversion at 80-90°C characteristic of enzymes showing low-temperature inactivation (LTI). (ulster.ac.uk)
  • He also solved early structures of the thermophilic bacterial enzyme thermolysin, the helix-turn-helix DNA-binding transcription factor lambda Cro repressor, and the light-antenna bacteriochlorophyll protein. (wikipedia.org)
  • The enzyme thermolysin catalyses the break up of peptides - components of proteins - in bacterial cells. (newscientist.com)
  • To compare the action of trypsin and thermolysin in the keratinocyte isolation using newborn foreskin . (bvsalud.org)
  • The number of colonies in the thermolysin group was significantly greater than in the trypsin group. (bvsalud.org)
  • B) Analysis of conformational changes due to thermolysin and trypsin. (asm.org)
  • srr7 virions were incubated with either 40 nM soMHVR (+) or BSA (−) at 37°C for 30 min, and then samples were digested with 15 μg of thermolysin/ml or 100 μg of trypsin/ml at 4°C for 30 min. (asm.org)
  • Although no atomic structure is available for this enzyme, site-directed mutagenesis studies have shown that its active site resembles closely that of the bacterial zinc-endopeptidase, thermolysin (EC 3.4.24.27). (inserm.fr)
  • Thermolysins constitute a family of secreted bacterial metalloproteases expressed, among others, by several pathogens. (edu.pl)
  • A volumetric yield of 46 mg thermolysin/L of bacterial culture and a yield of near 100% in relation to the total thermolysin present in TL-IBs were obtained. (butantan.gov.br)
  • Bacterial Thermolysin (TLN). (biolegend.com)
  • These enzymes are also termed 'neutral' proteinases or thermolysin -like proteinases (TLPs). (wikipedia.org)
  • The results, coupled with a molecular modeling study, suggest that Arg-717 of neprilysin corresponds to Arg-203 of thermolysin and that in both enzymes a hydrogen bond network exists, involving His-142, Asp-170, and Arg-203 in thermolysin and His-583, Asp-650, and Arg-717 in neprilysin, which is crucial for hydrolytic activity. (inserm.fr)
  • Enzymes from the thermolysin family are crucial factors in the pathogenesis of several diseases caused by bacteria and are potential targets for therapeutic interventions. (butantan.gov.br)
  • Enzymes that may be species variants of thermolysin are reported from Micrococcus caseolyticus [4] and Aspergillus oryzae [5]. (genome.jp)
  • S. Blumberg, Z. Vogel, and M. Altstein, Inhibition of enkephalin-degrading enzymes from rat brain and of thermolysin by amino acid hydroxamates, Life Sci. (springer.com)
  • In contrast to many proteins that undergo conformational changes upon heating and denaturation, thermolysin does not undergo any major conformational changes until at least 70 °C. The thermal stability of members of the TLP family is measured in terms of a T50 temperature. (wikipedia.org)
  • Preventing this calcium from originally binding to the molecule by mutation of its binding site, reduced thermolysin stability by 7 °C. However, while calcium binding makes a significant contribution to stabilising thermolysin, more crucial to stability is a small cluster of N-terminal domain amino acids located at the proteins surface. (wikipedia.org)
  • Thermolysin encoded by the gene LIC13322 of the causative agent of leptospirosis, Leptospira interrogans, was shown to cleave proteins from the Complement System. (butantan.gov.br)
  • Major proteins of these globules, and those treated with thermolysin after isolation, were identical. (springer.com)
  • However, the purified proteins were sensitive to thermolysin, indicating that domains of them are normally hidden in the globule matrix. (springer.com)
  • These results indicate that the five thermolysin-resistant proteins (apparent M r values: 56, 52, 32, 29, 27 kDa) are close to the surface and might be crucial for globule stabilization, whereas the thermolysin-accessible proteins are probably involved in globule/globule interactions and/or globule/eyespot-membrane interactions. (springer.com)
  • An analysis of successful SAD phasing on three proteins, lysozyme, glucose isomerase and thermolysin based on the signal of weak anomalous scatterers such as sulfur atom and chloride ion have been carried out. (scirp.org)
  • iv) Fewer detected peptides from thermolysin shaving originate from the transmembrane regions. (diva-portal.org)
  • Novel Angiotensin I-Converting Enzyme Inhibitory Peptides Found in a Thermolysin-Treated Elastin with Antihypertensive Activity. (peptide.com)
  • After digestion by thermolysin and pronase, cofactor peptides were purified by HPLC and sequenced. (sigmaaldrich.com)
  • Crystal structure analyses of thermolysin in complex with its inhibitors. (expasy.org)
  • The ability of comparative molecular field analysis (CoMFA), a three-dimensional, quantitative structure-activity relationship (3-D QSAR) paradigm, to predict the activity of inhibitors of angiotensin-converting enzyme (ACE) and thermolysin was examined. (wustl.edu)
  • 1987, 1, 3-16) were comparable to those derived for the thermolysin inhibitors using alignment rules defined by crystallographic data. (wustl.edu)
  • Experimentally derived alignment rules based on known structures of three-dimensional complexes produced predictive correlations for thermolysin inhibitors comparable, but not superior, to the correlations for ACE inhibitors based on alignment rules which were computationally deduced. (wustl.edu)
  • Methyl, Ethyl, Propyl, Butyl: Futile But Not for Water, as the Correlation of Structure and Thermodynamic Signature Shows in a Congeneric Series of Thermolysin Inhibitors. (cathdb.info)
  • Circular dichroism and fluorescence studies of thermolysin in the same solvent mixtures reveal discontinuous changes in the overall secondary and tertiary protein structure that correlate well with the reversible differences in catalytic activity. (strath.ac.uk)
  • Price, N.C. / Reversible acetonitrile-induced inactivation/activation of thermolysin . (strath.ac.uk)
  • Inouye, K , Lee, SB & Tonomura, B 1996, ' Effect of amino acid residues at the cleavable site of substrates on the remarkable activation of thermolysin by salts ', Biochemical Journal , vol. 315, no. 1, pp. 133-138. (elsevier.com)
  • Several species including Vibrio cholerae and V. vulnificus are known to produce thermolysin-like metalloproteases termed vibriolysin. (frontiersin.org)
  • In the present study, recombinant PAE was purified and compared with thermolysin in different organic solvents. (magiran.com)
  • However, the production of this recombinant protein using traditional refolding processes with high levels of denaturing reagents for thermolysin inclusion bodies (TL-IBs) solubilization results in poor recovery and low proteolytic activity probably due to improper refolding of the protein. (butantan.gov.br)
  • Thermolysin specifically catalyzes the hydrolysis of peptide bonds containing hydrophobic amino acids. (wikipedia.org)
  • However thermolysin is also widely used for peptide bond formation through the reverse reaction of hydrolysis. (wikipedia.org)
  • Based on the numerous homologies that deformylase displays with thermolysin and metzincins, a mechanism of enzyme:substrate recognition and hydrolysis could finally be proposed. (nih.gov)
  • Evidence by site-directed mutagenesis that arginine 203 of thermolysin and arginine 717 of neprilysin (neutral endopeptidase) play equivalent critical roles in substrate hydrolysis and inhibitor binding. (inserm.fr)
  • The activity of thermolysin in the hydrolysis of N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide and N-carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester is remarkably enhanced in the presence of high concentrations (1-5 M) of neutral salts. (elsevier.com)
  • However, activity of thermolysin was abolished in all organic solvents. (magiran.com)
  • Studies on the contribution of calcium to thermolysin stability have shown that upon thermal inactivation a single calcium ion is released from the molecule. (wikipedia.org)
  • Dette er en interessant observasjon siden man antar at spesifikke hemmere må binde til bindingsseter som ikke inkulderer det katalytiske Zinc atomet, siden katalytisk Zinc finnes i alle zinc metalloproteaser. (uit.no)
  • The predictive ability of the ACE model for a series of molecules not included in the training set was improved by the addition of a zinc indicator variable which explicitly defined the nature of the zinc-ligand interaction, an effect not observed within the thermolysin series. (wustl.edu)
  • Thermolysin: a zinc metalloenzyme. (genome.jp)
  • W. Structural basis of the action of thermolysin and related zinc peptidases. (genome.jp)
  • The overall topology of this domain is shared by the archetypal zinc-endopeptidase thermolysin. (ebi.ac.uk)
  • Thermolysin showed a biphasic log k versus 1/T plot consistent with the two-state model for protein and enzyme denaturation. (ulster.ac.uk)
  • This group of metallopeptidases belong to the MEROPS peptidase family M12, subfamily M12A (astacin family, clan MA(M)). The protein fold of the peptidase domain for members of this family resembles that of thermolysin, the type example for clan MA and the predicted active site residues for members of this family and thermolysin occur in the motif HEXXH [ PMID: 7674922 ]. (ebi.ac.uk)
  • In particular a phenylalanine (F) at amino acid position 63 and a proline (P) at amino acid position 69 contribute significantly to thermolysin stability. (wikipedia.org)
  • Amino-acid sequence of thermolysin. (genome.jp)
  • Human NEP can be classified as a thermolysin-type metalloendopeptidase (2,10,11). (springer.com)
  • One active site residue of thermolysin, Arg-203, is involved in inhibitor binding by forming hydrogen bonds with the carbonyl group of a residue in the P1 position and also participates in a hydrogen bond network involving Asp-170. (inserm.fr)
  • Experimental MAD phased structure of thermolysin in complex with inhibitor JC65. (expasy.org)
  • Sequence alignment data shows that Arg-717 of neprilysin could play a similar role to Arg-203 of thermolysin. (inserm.fr)
  • Thermolysin is catalytically inactive in mixtures or 70-15% acetonitrile in aqueous buffer. (strath.ac.uk)
  • The kinetics of the thermolysin-catalyzed synthesis of N-(benzyloxycarbonyl)-L-phenylalanyl-L-leucine ethyl ester (Z-Phe-LeuOEt) from N-(benzyloxycarbonyl)-L-phenylalanine (Z-Phe) and L-leucine ethyl ester (LeuOEt) in an ethyl acetate saturated aqueous system in a batch operation were studied. (elsevier.com)
  • This was investigated by site-directed mutagenesis with Arg-203 of thermolysin and Arg-717 of neprilysin being replaced by methionine residues. (inserm.fr)
  • Within the pH and temperature range examined, the k cat value for the synthesis of Z-Phe-LeuOEt reached a maximum at pH 7.0 and 45°C, whereas the affinity between Z-Phe and thermolysin reached a maximum at pH 6.0 and 40°C. The inhibitory effect of Z-Phe on the condensation reaction decreased as the pH and temperature decreased. (elsevier.com)
  • Crystal structures of alpha-mercaptoacyldipeptides in the thermolysin active site: structural parameters for a Zn monodentation or bidentation in metalloendopeptidases. (cathdb.info)
  • Thermolysin is synthesized as a pre-proenzyme consisting of a signal peptide 28 amino acids long, a pro-peptide 204 amino acids long and the mature enzyme itself 316 amino acids in length. (wikipedia.org)
  • Human FXI (hFXI) activity is measured by its ability to cleave a fluorogenic peptide substrate Boc-IEGR-AMC after hFXI is activated by thermolysin. (biolegend.com)
  • SEC-purified thermolysin suffers fragmentation, likely due to autoproteolysis and presents proteolytic activity against complement C3 a-chain, possibly by a generation of a C3b-like molecule. (butantan.gov.br)
  • Buchanan JD, Corbett RJ, Roche RS (1986) The thermodynamics of calcium binding to thermolysin. (edu.pl)
  • The proteolytic activity of thermolysin against C3 was time and dose-dependent. (butantan.gov.br)
  • Thermolysin-activated human FXI hydrolyzes a fluorogenic substrate, Boc-IEGR-AMC, with a specific activity value greater than 200 pmol/µg/min. (biolegend.com)
  • Thermolysin has a T50 value of 86.9 °C, making it the most thermo stable member of the TLP family. (wikipedia.org)
  • The internal region contains the HEXXH catalytic domain that is conserved in members of the thermolysin family. (edu.pl)
  • These globules were not stable and easily fused, whereas thermolysin-treated globules were relatively stable. (springer.com)
  • Thermolysin was added to improve the yield of epithelial cells , while endothelin-3 was added to stimulate their growth . (bvsalud.org)
  • Using thermolysin shaving we can also provide a clear separation between the best and the less accurate topology predictors, indicating that using data from shaving can provide valuable information when developing new topology predictors. (diva-portal.org)
  • This led, in both cases, to decreases in kcat/Km values, of 122-fold for neprilysin and 2300-fold for thermolysin, essentially due to changes in kcat. (inserm.fr)
  • The highest concentration of 2- O -lauroyl sucrose observed was 23.7 mM after 24 hours in the thermolysin-catalysed reaction. (aau.dk)
  • The concentration of sucrose laurate was observed to decline with reaction times above 6 hours in the reaction with thermolysin due to oligoester formation, and declining concentration with increasing reaction time was also observed for some specific sucrose laurate regioisomers in the reactions with other enzyme formulations. (aau.dk)
  • This method used thermolysin as it selectively digests the dermo-epidermal junction. (bvsalud.org)
  • Differentiated human intestinal epithelial cells , with the expression of surface markers (cytokeratins 8, 18 and mouse intestinal mucosa -1/39) and secretion of cytokines (sucrase-isomaltase, aminopeptidase N and dipeptidylpeptidase IV), may be cultured by the thermolysin and endothelin-3 method and maintained for at least 20 passages. (bvsalud.org)