Hemorrhagic principles in the venom of Bitis arietans, a viperous snake. II. Enzymatic properties with special reference to substrate specificity.
The optimal pH of the proteinase activity of hemorrhagins, BHRa and BHRb, isolated from the venom of Bitis arietans (puff adder) is pH 9. The activity was inhibited by metal chelating agents such as EDTA, 1,10-phenanthroline and 8-hydroxyquinoline, but not by phenylmethanesulfonyl fluoride and soybean trypsin inhibitor, suggesting that they are metalloproteinases. The hemorrhagins hydrolyzed all gelatin preparations derived from types I, II, III and IV collagen. On the other hand, only type IV native collagen was hydrolyzed. Gel electrophoretic profiles of type IV collagen hydrolysates suggested that the hemorrhagins affect the collagen helical chains at different cleavage sites. The hemorrhagins hydrolyzed several synthetic peptides such as angiotensin I and luteinizing hormone-releasing hormone, but not synthetic substrates for bacterial and animal collagenases. The hydrolysis of various peptides indicated that the hemorrhagins are endopeptidases. The insulin B chain is cleaved by BHRa and BHRb at 11 and 10 positions, respectively. The substrate specificity of the hemorrhagins was compared with those of known hemorrhagic and nonhemorrhagic venom proteinases.
Yamakawa
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Y
Department of Biochemistry and Cell Biology, National Institute of Health, Tokyo, Japan.
Omori-Satoh
T
T
Mebs
D
D
D000595:Amino Acid Sequence; D001665:Binding Sites; D003094:Collagen / Q000378:metabolism; D010450:Endopeptidases / Q000378:metabolism; D005780:Gelatin / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007328:Insulin / Q000031:analogs & derivatives; D008969:Molecular Sequence Data; D011480:Protease Inhibitors / Q000378:metabolism; D030162:Reptilian Proteins; D013379:Substrate Specificity; D014757:Viper Venoms / Q000037:antagonists & inhibitors / Q000201:enzymology* / Q000302:isolation & purification
D007328:Insulin; D011480:Protease Inhibitors; D030162:Reptilian Proteins; D014757:Viper Venoms; C091389:hemorrhagin a, Bitis arietans; C091390:hemorrhagin b, Bitis arietans; D005780:Gelatin; D003094:Collagen; D010450:Endopeptidases