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. 4@silica 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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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 ...

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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 ...

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All text is © British Library Board and is available under a Creative Commons Attribution Licence, except where otherwise stated.. ...

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