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K Ohlsson I Olsson M Delshammar H Schiessler 《Hoppe-Seyler's Zeitschrift für physiologische Chemie》1976,357(9):1245-1250
The lysosome-like granules of human and canine granulocytes contain an enzyme with elastinolytic activity. The enzymatic behaviour of these elastases was further characterized using the protein substrates elastin-orcein and azocasein and the synthetic substrates tert.-butyloxycarbonyl-alanine p-nitrophenylester (Boc-Ala-ONp) and 3-carboxypropionyl-L-alanyl-L-alanyl-L-alanine p-nitroanilide (Suc-Ala3-NHNp) in photometric assays. The affinities of the granulocyte elastases and of porcine pancreatic elastase to these substrates are very similar, e.g. human granulocyte elastase: KM (Boc-Ala-ONp) = 0.35mM, KM (Suc-Ala3-NHNp) = 1.25mM, porcine pancreatic elastase: KM (Boc-Ala-ONp) = 0.3mM, KM (Suc-Ala3-NHNp) - 1.15mM. The most convenient substrate for the assay of human and dog granulocyte elastases and for kinetic measurements with these enzymes is Suc-Ala3-NHNp. Using this substrate, the dissociation constant of the complex of human granulocyte elastase with human alpha1-antitrypsin could be determined (Ki = 3.5 x 10(-10)M). 相似文献
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《Biochemical medicine》1982,27(2):143-153
Mixtures of human and dog pancreatic proteinases obtained from human pancreatic juice or extracts of canine pancreas were selectively inhibited using nine naturally occurring inhibitors of serine proteinases. Inhibition of trypsin, chymotrypsin, and elastase was monitored using specific synthetic substrates.Dog trypsin was inhibited by all nine inhibitors, while only four inhibitors significantly inhibited dog chymotrypsin and elastase. Inhibition spectra of human enzymes were completely different. Only three inhibitors exhibited significant inhibition of human trypsin. Human chymotrypsin and protease E (elastase) were inhibited only by 2 to 3 inhibitors and, in most cases, the amount of the inhibitor required to achieve full or 80% inhibition greatly exceeded the amount needed for the respective dog enzymes. Trasylol, the most widely used, commercially available inhibitor, inhibited both human and dog trypsins, but was totally ineffective against human and dog chymotrypsins and elastases. It was found that the most effective polyvalent inhibitors of human enzymes were chickpea and lima bean trypsin inhibitors. It was indicated by these results that strong species specificity exists and the invalidity of using animal models for the study of inhibition of human pancreatic proteinases was emphasized. 相似文献
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E L Gustavsson K Ohlsson A S Olsson 《Hoppe-Seyler's Zeitschrift für physiologische Chemie》1980,361(2):169-176
1 ml of human serum inhibits about 0.9 mg of purified human pancreatic elastase owing to complexation with alpha 1-antitrypsin and alpha 2-macroglobulin. On addition to serum, elastase is preferentially bound by alpha 2-macroglobulin. The complexes between elastase and alpha 1-antitrypsin and alpha 2-macroglobulin, respectively, migrate as alpha 2-globulin on agarose gel electrophoresis. Elastase bound by alpha 1-antitrypsin is precipitated by antibodies against enzyme as well as inhibitor, while the alpha 2-macroglobulin-bound elastase is only precipitated by antibodies against the inhibitor. The molar combining ratio for elastase/alpha 1-antitrypsin is 1:1 and for elastase/alpha 2-macroglobulin 2:1. The elastase bound by alpha 2-macroglobulin retains its activity against low molecular weight substrates, while that bound by alpha 1-antitrypsin is enzymologically inactive. 相似文献
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The human uterine estrogen receptor has a site which regulates estrogen binding and which structurally resembles the substrate binding site of chymotrypsin. The hormone binding capacity and the affinity of the receptor is decreased in the presence of 4 mM serine protease inhibitors tosyl-lysine chloromethyl ketone and diisopropylfluorophosphate and the protease substrates tryptophan methyl ester and toluene sulphonyl-arginine methyl ester. The protease inhibitors tosylamide-phenylethyl-chloromethyl ketone and phenyl methyl sulphonyl fluoride caused an increase in the binding capacity whereas the affinity was decreased. 相似文献
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Very long chain fatty acids (FAs) are important components of different classes of lipids in all organisms from bacteria to man. They include also, usually as minor components, odd-numbered FAs. These have so far been given little attention because of technical difficulties inherent in their detection and identification. Current modern analytical methods such as GC–MS and/or LC–MS make this detection and identification possible, and should promote a study of their properties. This review brings, in a concise manner, most of the currently available information about these FAs, their occurrence in different organisms, their structure and other properties. It should provide an impetus for further research into these very interesting compounds whose chemical, biochemical and biological properties are poorly known. 相似文献
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The interaction of four purified alpha-amylase (1,4-alpha-D-glucan glucanohydrolase, EC 3.2.1.1) inhibitors with human salivary and pancreatic alpha-amylases was investigated. The inhibitory activity of the four proteins towards salivary alpha-amylase was significantly increased by pre-incubation of the enzyme with inhibitor before adding substrate. This effect was not observed with the inhibition of pancreatic alpha-amylase by inhibitors 1 and 2. Inhibition of both amylases was affected to different degrees by incubating starch with inhibitor prior to the addition of enzyme. Maltose, at concentrations which only slightly affected amylase activity, prevented the inhibition of both enzymes by all four inhibitors. Gel filtration studies on salivary amylase-inhibitor mixtures showed the formation of EI complexes on a mol-to-mol ratio. A similar complex between pancreatic alpha-amylase and inhibitor 4 was observed, though complex formation between pancreatic alpha-amylase and the other inhibitors was not clearly demonstrated. 相似文献
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Classification of microbial, plant and animal cytolysins based on their membrane-damaging effects of human fibroblasts 总被引:9,自引:0,他引:9
38 cytolytic agents of mainly microbial origin were investigated with respect to membrane-damaging activity on human diploid fibroblasts. Increased plasma membrane permeability was measured as leakage of three defined cytoplasmic markers of various sizes: alpha-aminoisobutyric acid, uridine nucleotides and ribosomal RNA. The relative leakages of these markers, caused by different concentrations of the various cytolysins, yielded a leakage pattern for each substance. Five distinct types of leakage patterns were obtained. These were transformed into numerical expressions by calculating the ratios between the amounts of cytolysin needed to release 50% of the nucleotide and ribosomal RNA markers and the amounts required to release 50% of the alpha-aminoisobutyric acid marker (ED50 ratios). A classification of the cytolysins into five groups was arrived at on the basis of the different types of leakage patterns with the aid of reference cytolysins with well-known mechanisms of membrane interaction. These groups comprised: (1) detergent-like agents, (2) agents interacting with only certain constituents of the cell membrane, (3) agents interacting with specific receptor molecules in the membrane, (4) agents inducing small functional holes of a definable size, and (5) agents inducing only a very limited increase in plasma membrane permeability. The system may be useful for characterization and differentiation of new cytolytic agents of various sources as it divides membrane-damaging agents into separate groups on the basis of their principal function on intact human cells. 相似文献
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S C Alter J A Kramps A Janoff L B Schwartz 《Archives of biochemistry and biophysics》1990,276(1):26-31
Tryptase from human mast cells has been shown (in vitro) to catalyze the destruction of fibrinogen and high-molecular-weight kininogen as well as the activation of C3a and collagenase. Although large amounts of tryptase are released in tissues by degranulating mast cells and levels as high as 1000 ng/ml have been measured in the circulation following systemic anaphylaxis, no specific physiologic inhibitor has yet been found for the protease. The current work tests several more inhibitors for their effects on tryptase and examines any effect of tryptase on these inhibitors. First, antileukoprotease and low-molecular-weight elastase inhibitor from human lung and hirudin and antithrombin III had no effect on tryptase activity in vitro. Second, the possibility that tryptase, being insensitive to the effects of inhibitors, might instead destroy them was also considered. Tryptase failed to cleave and inactivate antileukoprotease, low-molecular-weight elastase inhibitor, alpha 1 protease inhibitor, alpha 2 macroglobulin, and antithrombin III. Third, based on the knowledge that tryptase stability is regulated by its interaction with heparin, antithrombin III was used as a model heparin-binding protein to demonstrate that a protein competitor for heparin-binding sites, presumably by displacement of tryptase, destabilizes this enzyme. Conversely, tryptase, in excess, blocked the binding of antithrombin III to heparin, thereby attenuating the heparin-mediated inhibition of thrombin by antithrombin III. 相似文献
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This review, issued in two parts, describes the information on the structure and biological activity of animal alkaloids derived from polymethyleneamines and produced by marine organisms, wasps, spiders, and microorganisms. Animal alkaloids are outstanding models for developing methods and drugs for the treatment of many human diseases. In the first part, we consider compounds produced by marine and microbial organisms. Some promising synthetic analogues of these alkaloids are used in developing modem preparations for the chelate therapy of excessive blood iron content and antituberculosis, antiproliferative, and immunosuppressive drugs. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2005, vol. 31, no. 6; see also http://www.maik.ru. 相似文献
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J A Knight R W Stephens G R Bushell P Ghosh T K Taylor 《Biochimica et biophysica acta》1979,584(2):304-310
Assays of several proteases, incorporating guanidinium chloride extracts of human femoral head cartilage and intervertebral disc, demonstrated that both tissues contain inhibitors of certain serine proteases. Trypsin, chymotrypsin and a granule extract of human polymorphonuclear leukocytes containing elastase and cathepsin G activities, were inhibited by low molecular weight fractions prepared by Sephadex G-75 chromatography. Using a radioassay, it was further shown that these fractions inhibit proteolysis of cartilage proteoglycan. The inhibitor in intervertebral disc is concentrated in the nucleus pulposus, with a decreasing gradient to the periphery of the annulus fibrosus. It is proposed that these inhibitors confer at least partial protection against pathological proteolysis of the proteoglycans in human articular cartilage and nucleus pulposus. 相似文献
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Two new double-headed protease inhibitors have been isolated from black-eyed peas. The isoinhibitors can be purified to homogeneity with greater than 90% recovery in a four-step procedure by means of sequential affinity chromatography on trypsin-Sepharose and chymotrypsin-Sepharose affinity columns. The isoinhibitors both have molecular weights near 8,000 and both have the same NH1-terminal residue serine. Black-eyed pea chymotrypsin and trypsin inhibitor (BEPCI) has an isoelectric point of 5.1 and inhibits trypsin and chymotrypsin simultaneously. Black-eyed pea trypsin inhibitor (BEPTI) has an isoelectric point of 6.5 and inhibits 2 molecules of trypsin simultaneously. BEPTI binds to chymotrypsin-Sepharose above pH 6 but does not inhibit chymotrypsin in the standard inhibitor assay with 10-3 M substrate. These new inhibitors are distinct from the Ventura inhibitor isolated from Serido black-eyed peas. An endogenous seed protease has been isolated from black-eyed peas by affinity chromatography on soybean inhibitor-carboxymethylcellulose affinity columns. A protease-BEPCI complex has been isolated by ion exchange chromatography. A dual physiological function of inhibition and protection of the seed protease is suggested as a plausible role of seed protease inhibitors. 相似文献