Pancreatic Serine Proteases Research Articles

Bovine factor X1 (Stuart factor): amino-acid sequence of heavey chain.

The amino-acid sequence of the heavy chain of bovine blood coagulation factor X1 (Stuart factor) isolated before and after activation has been determined. Sequence analysis was performed on fragments obtained by cleavage with cyanogen bromide and by tryptic digestion. Comparison of the complete sequence with those of other hepatic and pancreatic serine proteases demonstrates homology of the heavy chain of activated factor X1 (factor X1a) with the B chain of bovine thrombin as well as with bovine trypsin, chymotrypsins A and B, and porcine elastase. The activation peptide cleaved near the amino terminus by a protease from Russell's viper venom differs in both size and sequence from those of other serine proteases. With three exceptions, all of the residues which are important in the catalytic functions of trypsin and chymotrypsin occur in corresponding loci in the heavy chain of factor Xa. These finding suggest that the three-dimensional structure of the heavy chain is similar to that of the pancreatic serine proteases and that these enzymes have evolved from a common ancestral gene.

The phylogeny of trypsin-related serine proteases and their zymogens. New methods for the investigation of distant evolutionary relationships

The sequence of all presently known trypsin-related serine proteases and their zymogens of animal and bacterial origin were optimally aligned on the basis of three different scoring schemes for amino acid comparisons. Sequence homology was found to extend into the activation peptides. The gaps resulting from the alignment of the sequences of the active enzymes formed the basis for a new procedure based on position and number of gaps, which allowed the correct topology of the evolutionary relationship of thrombin and the pancreatic enzymes trypsin, chymotrypsin and elastase to be determined. The procedure was applied in an analogous manner to changes in disulfide bridges as well as to a selected set of amino acid positions. Evolutionary distances between proteins were estimated by minimum, base differences as well as according to the stochastic model of evolution. These distances were used successfully to find the best topology of evolutionary relationships. The fact that the branch lengths in evolutionary trees were less affected by the number of sequences considered when evolutionary distances between contemporary sequences were measured in minimum base differences than when measured according to the stochastic model of evolution, suggested in our specific case, that minimum base differences yielded estimates of evolutionary distance closer to reality than the stochastic model of evolution. All these techniques combined yielded the following picture for the evolution of the four protease families. Prothrombin and the zymogens of the pancreatic serine proteases had a common ancestor with tryptic specificity. After the initial divergence, the gene for trypsinogen duplicated. Evidence was found that the duplicated gene underwent drastic changes for a short period of time to become eventually the common ancestor of chymotrypsin and elastase. The phylogenetic tree elaborated for these enzyme families and the methods introduced to determine its topology, should readily allow determination of the attachment site of branches leading to newly sequenced serine proteases, provided their amino acid sequence can be aligned fairly unambiguously. In addition, the consequences of the alignment of the different serine proteases for the relationship of zymogen to enzyme are discussed.

Effect of pH on the catalytic activity of Streptomyces griseus protease 3.

The kinetics of the Streptomyces griseus protease-3-catalyzed hydrolysis of the non-specific ester substrate p-nitrophenylacetate and the specific peptide substrate glutaryl-l-phenylalanine-p-nitroanilide are consistent with a three-step mechanism of Michaelis complex formation followed by enzyme acylation and deacylation. Both substrates pre-equilibrate rapidly with the free enzyme, the acylation (deacylation) step being rate-limiting in the enzymatic hydrolysis of glutaryl-l-phenylalanine-p-nitroanilide (p-nitrophenylacetate). Examination of the effect of pH on steady-state and transient-state kinetic parameters for the enzymatic hydrolysis of the two substrates provides evidence that the stability of the Michaelis complex is essentially unaffected by pH over the range investigated, whereas both acylation and deacylation of the enzyme is dependent upon an ionizing group in the protein with apparent pKa of 6.6. It is concluded that closely similar mechanisms of activation of the reactive seryl residue may be operative in Streptomyces griseus protease 3 and pancreatic serine proteases such as chymotrypsin and elastase.

Isolation of alkaline proteinases from Aspergillus oryzae by one-step affinity chromatography on ovoinhibitor-sepharose column

1. 1. Alkaline proteinases from a crude preparation of Aspergillus oryzae were isolated by one-step affinity chromatography on a chicken ovoinhibitor-Sepharose column. The yield of the activity was 55% and the purified fraction had about 300-fold higher specific activity than the starting material. 2. 2. The purified fraction was found to be homogeneous in size but consisted of at least four electrophoretically distinct bands. The amino acid composition was estimated and found to be very similar to other known alkaline proteinases from Aspergillus species, thus indicating a strong genetic relationship. 3. 3. The esterolytic activity of the purified fraction was tested on specific esteratic substrates of pancreatic serine proteases. The purified fraction was active on all 3 substrates, thus indicating a rather broad specificity and the kinetic parameters were almost identical to those of pure alkaline proteinase from Aspergillus sojae. Like the latter, the purified fraction had an extremely high k cat on acetyl tri- l-alanine methyl ester.

Proteolysis in formation of a myocardial depressant factor during shock

ARTICLESProteolysis in formation of a myocardial depressant factor during shockJN Leffler, Y Litvin, Y Barenholz, and AM LeferJN Leffler, Y Litvin, Y Barenholz, and AM LeferPublished Online:01 Apr 1973https://doi.org/10.1152/ajplegacy.1973.224.4.824MoreSectionsPDF (2 MB)Download PDF ToolsExport citationAdd to favoritesGet permissionsTrack citations ShareShare onFacebookTwitterLinkedInEmailWeChat Previous Back to Top Next Download PDF FiguresReferencesRelatedInformationCited ByPancreatic Digestive Enzyme Blockade in the Intestine Increases Survival After Experimental ShockScience Translational Medicine, Vol. 5, No. 169Role of pancreatic enzymes in the development of multiple organ failure after shock11 July 2009 | Journal of Organ Dysfunction, Vol. 4, No. 3INTRALUMINAL PANCREATIC SERINE PROTEASE ACTIVITY, MUCOSAL PERMEABILITY, AND SHOCKShock, Vol. 26, No. 1Characterization of Two Classes of Pancreatic Shock Factors: Functional Differences Exhibited by Hydrophilic and Hydrophobic Shock FactorsShock, Vol. 20, No. 4Delayed dilated cardiomyopathy for major burn injuriesBurns, Vol. 29, No. 4Pancreatic Enzymes and Microvascular Cell Activation in Multiorgan FailureMicrocirculation, Vol. 8, No. 1The Pancreas as a Source of Cardiovascular Cell Activating FactorsMicrocirculation, Vol. 7, No. 3Plasma cathepsin activity and the role of hypovolemic shock in dehydrational death in the toad, bufo marinusComparative Biochemistry and Physiology Part A: Physiology, Vol. 90, No. 1Myocardial Depressant Substances in Septic ShockPain Control in Mass CasualtiesBeneficial action of naloxone in splanchnic artery occlusion shockExperientia, Vol. 37, No. 4A Study of the Myocardial Depressant Factor and Its Relative Influence in Drug/Alcohol Mortality1 July 1980 | Journal of Forensic Sciences, Vol. 25, No. 3Effect of suppression of endocrine or exocrine pancreatic function in hemorrhagic shockExperientia, Vol. 35, No. 1Beneficial action of new angiotensin-converting enzyme inhibitor (SQ 14,225) in hemorrhagic shock in cats.Circulation Research, Vol. 43, No. 4Isolation of myocardial depressant factor from plasma of dogs in hemorrhagic shockBiochimica et Biophysica Acta (BBA) - Protein Structure, Vol. 491, No. 1Effect of induced thrombocytopenia on experimental circulatory shockJournal of Surgical Research, Vol. 21, No. 6Myocardial depressant factor and circulatory shockKlinische Wochenschrift, Vol. 52, No. 8Factors influencing the in vitro production of a myocardial depressant factorBiochemical Medicine, Vol. 8, No. 2 More from this issue > Volume 224Issue 4April 1973Pages 824-831 Copyright & PermissionsCopyright © 1973 by American Physiological Societyhttps://doi.org/10.1152/ajplegacy.1973.224.4.824PubMed4698799History Published online 1 April 1973 Published in print 1 April 1973 Metrics

Pancreatic hydrolases and the formation of a myocardial depressant factor in shock.

Pancreatic hydrolases and the formation of a myocardial depressant factor in shock.

Amino-acid composition of crayfish trypsin.

THE homology in primary structure of mammalian pancreatic serine proteases is well established1,2. Progress has now been made in the elucidation of the evolutionary history of this group of proteases. Neurath and his co-workers3 studied the amino-acid composition of trypsinogen isolated from the pancreas of the spiny Pacific dogfish and presented a preliminary examination of the primary structure of this enzyme. From these data a high degree of homology as compared with bovine trypsinogen can be inferred. It is now of interest what will be found beyond the gap that separates vertebrate and invertebrate animals. Invertebrate proteases are difficult to investigate because of the small amounts of material obtainable from these animals. In the crayfish Astacus leptodactylus we found an enzyme resembling mammalian trypsin in many respects4,5. It exhibits a distinct tryptic cleavage specificity when tried on the oxidized B-chain of insulin and is inhibited by all natural and synthetic trypsin inhibitors including TLCK. It is a serine protease and possesses the sequence –Asp–Ser–Gly– around the active centre serine residue6. In addition, extended immunological studies have been carried out on the crayfish trypsins of three species which revealed that they do occur as iso-enzymes varying in number and electrophoretic mobility7.

Primary structure of alpha-lytic protease: a bacterial homologue of the pancreatic serine proteases.

Abstract : Comparison of the amino-acid sequence of the bacterial alpha-lytic protease wit the mammalian pancreatic proteases shows homologies in the regions of the N-terminus, the single histidine, an aspartic acid residue, the active serine, and in the positions of the three disulphide bridges. The partial sequence of a trypsin-like enzyme from Streptomyces griseus provides further evidence for divergent evolution of this class of enzymes from a common ancestral gene. (Author)

Inactivation of bovine thrombin by nitrous acid.

Bovine thrombin is rapidly inactivated by nitrous acid at pH 4.35 and 0°. The clotting activity falls off more rapidly than the activity towards N-benzoylarginine ethyl ester. The loss of esterase activity parallels the loss of the N-terminal isoleucine residue. N-Benzoylarginine ethyl ester gives almost full protection of the esterase activity and the N-terminal isoleucine, but only partial protection of the clotting activity. These results suggest strongly that this residue is essential for activity and indicate that thrombin resembles the pancreatic serine proteinases in this respect. The initial rate of inactivation of thrombin by nitrous acid is higher than that of trypsin, and considerably higher than that of elastase and α-chymotrypsin under the same conditions, suggesting that the N-terminal residue is more readily accessible to the reagent. The experiments strengthen the hypothesis that all mechanisms of prothrombin activation are proteolytic and suggest that they involve the liberation of the essential N-terminal isoleucine.

Crystal Data for a Bacterial Serine Protease

α-LYTIC protease is one of several proteolytic enzymes isolated from culture filtrates of a soil bacillus Sorangium sp. (Myxobacter 405)1. Chemical and kinetic studies of this enzyme have shown many remarkable homologies with the pancreatic serine proteases chymotrypsins A and B, trypsin and elastase. For example, α-lytic protease has exactly the same sequence (.Gly.Asp.Ser*.Gly.Gly.) around the active serine as have the pancreatic enzymes1,2. Moreover, the sequence around the single histidine residue in the enzyme (Val.Thr.Ala.Gly.His.Cys.Gly.)3 closely matches the sequence around His 57 of chymotrypsins A4,5 and B6, and the similar sequences around the corresponding histidine residues of trypsin7,8 and pancreatic elastase9.

Amino acid sequence around the histidine residue of the alpha-lytic protease of Sorangium sp., a bacterial homolog of the pancreatic serine proteases.

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTAmino acid sequence around the histidine residue of the .alpha.-lytic protease of Sorangium species, a bacterial homolog of the pancreatic serine proteasesLawrence B. Smillie and Donald R. WhitakerCite this: J. Am. Chem. Soc. 1967, 89, 13, 3350–3352Publication Date (Print):June 1, 1967Publication History Published online1 May 2002Published inissue 1 June 1967https://doi.org/10.1021/ja00989a046RIGHTS & PERMISSIONSArticle Views34Altmetric-Citations23LEARN ABOUT THESE METRICSArticle Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated. Share Add toView InAdd Full Text with ReferenceAdd Description ExportRISCitationCitation and abstractCitation and referencesMore Options Share onFacebookTwitterWechatLinked InReddit PDF (352 KB) Get e-Alerts Get e-Alerts