Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.4.21.7 (plasmin)
 9,023 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The scientific literature on milk proteases, along with recent findings in the author's laboratory, are summarized and reviewed comprehensively. Emphasis is on detection of proteolytic enzymes and their activity, purification and kinetic characterization of the isolated enzymes, and technological problems associated with proteolytic enzymes in milk and milk products. Two serine proteinases isolated from milk are compared with plasmin of bovine blood serum. Results from these comparisons strongly suggest that milk proteinase I and plasmin are identical. Proteolysis studies with cold stored milk indicate a direct relationship between gamma-casein formation and milk proteinase association with casein micelles.
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PMID:New aspects of naturally occurring proteases in bovine milk. 622 88

It has been previously demonstrated that commercial bacterial fibrinolysin (EC 3.4.21.7) selectively cleaves the bond between Met-53 and Ala-54 in ovine prolactin (199 amino acids). A one-step purification procedure on DEAE-cellulose for Protease F, which is the active component of bacterial fibrinolysin, and properties of the purified enzyme are reported. The enzyme is homogeneous as judged by acrylamide gel electrophoresis. Its molecular weight, calculated from gel filtration experiments on Sephadex G-100, is around 13,800. Amino acid analyses do not reveal the presence of any half-cystines. The presence of one tryptophan residue per enzyme molecule was resolved from the fluorescence spectrum. Amino terminal analysis showed that leucine was at the amino terminal position. Protease F hydrolyzes casein and synthetic specific substrates for chymotrypsin and elastase esterases but not for trypsin esterases. It is fully inhibited by phenylmethylsulfonyl fluoride, by chicken ovoinhibitor, and by Chymotrypsin Inhibitor I from potatoes but not by the trypsin-chymotrypsin inhibitors from soybeans and chick peas or by tosyl-L-phenylalanine chloromethyl ketone. The enzyme is stable at room temperature and in the cold, it is not affected by dialysis or by freezing and thawing, but it is inactivated during freeze-drying. The circular dichroism spectra of Protease F indicate an approximate 20% alpha-helix content of the enzyme with a considerable similarity to those of subtilisin, elastase, and beta-trypsin. The relatively low molecular weight of Protease F, the absence of intrachain disulfide bridges, and the fact that it is inhibited by several, but not all, chymotrypsin inhibitors suggest that it may differ phylogenetically from the known serine proteases.
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PMID:Purification and properties of protease F, a bacterial enzyme with chymotrypsin and elastase specificities. 622 44

Growth of Streptococcus thermophilus was faster in mastitic milk, aseptically collected following intramammary infusion of Escherichia coli endotoxin, than in normal aseptically collected milk. The contribution of polymorphonuclear leucocytes (PMN), of plasma and plasmin to this stimulation has been investigated. Addition of plasmin to low cell count milk stimulated growth of the streptococcus to the same degree as mastitic milk. However, addition of plasma or PMN from blood or from milk were less stimulatory. The part played by these components in providing casein breakdown products for the weakly proteolytic Str. thermophilus in mastitic milk is discussed.
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PMID:Stimulation of Streptococcus thermophilus growth in mastitic milk. 623 18

A proteinase which can activate human, dog and rat plasminogen to plasmin has been isolated from the urine of female rats, using affinity chromatography on benzamidine-coupled Sepharose. Inhibition by diisopropylfluorophosphate, tosyl-L-lysine chloromethylketone and benzamidine classified the enzyme as trypsin-like. The proteinase has weak activity on alpha-casein and hemoglobin, but will not lyse fibrin clots. It readily cleaves arginyl amides, including synthetic substrates specific for human glandular kallikrein and other serine proteinases. A chromogenic substrate for human urokinase (pyro Glu-Gly-Arg-pNA) is a poor substrate for the rat proteinase. Characteristics of the enzyme, such as its molecular weight (25 900), kinetic parameters and inhibition by aprotinin, indicate that this proteinase is esterase A, described by several investigators. Esterase A is shown not to be a true urinary plasminogen activator but rather is a unique arginine-specific proteinase. Urokinase-like and kallikrein-like activity are part of a broader proteolytic activity displayed by this enzyme.
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PMID:Esterase A is a proteinase from rat urine that can activate plasminogen. 623 43

Plasmin cleaves isolated human beta-casein to form specific fragments in a manner similar to the generation of gamma 1-, gamma 2-, and gamma 3-caseins from the bovine homologue. Identification of a protein previously isolated from human milk as a specific plasmin cleaved portion of beta-casein indicates that endogenous plasmin is active in whole milk. These findings suggest that protease activity should be considered in casein quantitation or isolation of components from human milk.
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PMID:Plasmin cleaves human beta-casein. 624 Feb 66

Certain strains of Moraxella bovis produce tissue-damaging enzymes which may initiate or potentiate infectious bovine keratoconjunctivitis. Thirteen reference strains of this species were characterized physiologically and screened for production of various enzymes by some conventional biochemical tests and the API ZYM system (Analytab Products, Plainview, N.Y.). All 13 strains were hemolytic. All hydrolyzed Tween 80 and Tween 85 and displayed C4 esterase, C8 esterase-lipase, and C14 lipase activities. All produced phosphoamidase and phosphatase. All were able to hydrolyze casein and gelatin. All produced leucine and valine aminopeptidases and fibrinolysin. Twelve produced hyaluronidase or were agarolytic. Three hydrolyzed chondroitin sulfate. Nine strains autoagglutinated. Five produced catalase, and two produced cystine aminopeptidase.
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PMID:Hydrolytic enzymes of Moraxella bovis. 625 99

Native proteolytic enzymes in good quality normal bovine milk readily hydrolysed the caseins during incubation or storage, producing the gamma-caseins, proteose-peptone components 5 (PP5) and 8-fast (PP8F) and a considerable number of other unidentified fragments, many of which were also subsequently found in the proteose-peptone fraction. The rate of casein hydrolysis was greater in pasteurized than in raw milk, with beta-casein being slightly more susceptible to attack than alpha S1-casein. Measurements of gamma-casein and proteose-peptone formation have been made and it was found that PP5 was an intermediate product that was subject to further proteolysis while PP8F was a stable end-product. With the exception of component 3 (PP3), virtually all constituents of the proteose-peptone fraction increased during storage and appeared to be products of the action of proteolytic enzymes. Further evidence was obtained from the effects of various inhibitors that the principal proteinase of normal milk is plasmin, but slight differences were apparent between the protein breakdown patterns induced by storage and by added plasmin, which was consistent with the presence of more than one proteinase. Incubations in the presence of soya bean trypsin inhibitor to prevent plasmin action clearly revealed that another enzyme(s) was also involved.
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PMID:Proteinases in normal bovine milk and their action on caseins. 634 22

Electrophoretic analysis of plasminogen activators from pig heart, human uterus, human plasma and human melanoma cells was performed in SDS-polyacrylamide gradient slab gels containing plasminogen and casein. Direct visualization of activator activity bands in polyacrylamide gels was achieved after removal of SDS, incubation in buffer, and staining with Coomassie brilliant blue. Tissue activator extracted from pig hearts displayed a molecular weight of 72000 and migrated similarly to activator secreted by human melanoma cells and to one activator component present in extracts of human uterus. Immunoadsorption experiments with melanoma cell activator antiserum indicated that these 72-kDa activators are all related immunologically. Human uterus also contained a second activator component with a molecular weight 55000, which migrated similarly to a higher molecular weight component of urokinase and cross-reacted with urokinase antiserum. We conclude that the 72-kDa uterine activator component represents a tissue activator and the 55-kDa component represents a urokinase-like activator. A euglobulin solution from venous occlusion plasma displayed multiple bands of plasmin activity in the Mr range 85000-96000. Two activator components were also present, one of Mr 72000 and another of Mr 62000. The 72-kDa euglobulin activator was adsorbed by MCA antiserum, and we conclude that this component represents vascular activator. The 62000 activator also had weak plasminogen-independent caseinolytic activity and was not affected by either melanoma cell activator or urokinase antisera. Conclusions concerning its identity cannot be made at this time.
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PMID:Comparative electrophoretic analysis of human and porcine plasminogen activators in SDS-polyacrylamide gels containing plasminogen and casein. 634 80

The alteration of human and porcine plasmin and the influence of EACA and AMCHA on their activity were investigated. Solutions of both plasmins undergo storage induced alteration, which is best recorded by Chromozym PL, whereas the other chromogenic substrates, S-2251 and S-2302, and casein are less sensitive, and the fibrin plate inadequate. Plasmin amidolytic and fibrinolytic activity is maximally enhanced at 7.6 x 10(-3) M EACA and 6.4 x 10(-4) M AMCHA, and decay through storage is reversed. The caseinolytic activity seems slightly inhibited at the same EACA and AMCHA concentrations. Our results show: 1. The quotient: plasmin activity towards Chromozym PL/Activity towards S-2251 is a useful indicator of "plasmin quality". The quotient decreases markedly upon storage of plasmin solutions. 2. Plasmin stability is improved in the presence of AMCHA. 3. It is valueless to add EACA or AMCHA to inhibit plasmin in amidolytic assays since the chosen concentration enhances the amidolytic activity of already formed plasmin.
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PMID:Different assessment of plasmin with different substrates. In vitro alteration of plasmin, influence of epsilon-aminocaproic acid and tranexamic acid upon its activity. 645 Apr 67

A method was investigated for the sensitive radiochemical assay of milk protein transformations. beta-Casein was reductively methylated with NaBH4 and H14CHO giving a product of spec. act. 4.42 microCi/mg in which a maximum of 12% of the lysine residues were labelled. Methylation did not alter the electrophoretic or chromatographic properties of the protein, or its pattern of proteolysis by plasmin. The substrate susceptibility of 14C-methylated beta-casein towards plasmin was determined by measuring radioactivity transfer to the proteolytic fragments gamma 2- and gamma 3-casein. Compared with the native protein, no decrease in substrate susceptibility was detected. The presence in milk of the plasmin-like enzyme, milk proteinase, was demonstrated and its activity at 4 degrees C was quantified by examination of the fragmentation pattern of added 14C-methylated beta-casein. It was concluded that 14C-methylated protein substrates, prepared by reductive methylation, are well suited to the study of hydrolytic enzymes and that they can provide valuable information on milk protein transformations. In particular, no interference was encountered with the rate of cleavage by plasmin when the level of methylation was kept to a minimum.
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PMID:14C-Methylated beta-casein as a substrate for plasmin, and its application to the study of milk protein transformations. 645 Jun 17


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