Characterization of hemorrhagic principles from Trimeresurus gramineus snake venom.
In addition to alpha-fibrinogenase (hemorrhagin I, HR1), a potent hemorrhagic principle (hemorrhagin II, HR2) was purified from Trimeresurus gramineus venom. It was homogeneous as judged by SDS-polyacrylamide gel electrophoresis. HR2 was a single peptide chain containing 10% carbohydrate with a molecular weight of 81,500. It possessed 669 amino acid residues per molecule, while HR1 contained only 203 amino acid residues per molecule with a molecular weight of 23,500. Both hemorrhagins possessed proteolytic activities toward fibrinogen, casein and azocoll. However, the proteolytic activities of HR1 were much more potent than those of HR2. They were devoid of TAME-esterase and phospholipase A2 activities which were found in crude venom. beta-Mercaptoethanol and antivenin completely inhibited the hemorrhagic activities of HR1 and HR2, while epsilon-aminocaproic acid, trasylol, p-bromophenacyl bromide, phenylmethanesulfonyl fluoride and soybean trypsin inhibitor did not. EDTA completely inhibited the hemorrhagic, fibrinogenolytic and caseinolytic activities of HR1. EDTA also completely inhibited the caseinolytic and fibrinogenolytic activities of HR2, but only partially inhibited its hemorrhagic activity. Subsequent addition of Zn2+ (5 mM) reversed the EDTA-induced inhibitory effect on the hemorrhagic activity of HR1. However, ZN2+ did not reverse the EDTA-induced inhibitory effect on the HR2-induced hemorrhagic activity. These hemorrhagins were found to be ZN2+-containing metalloproteinases. Therefore, the hemorrhagic activity of HR1 seems to be related to its proteolytic activity while that of HR2 seems to be unrelated to its proteolytic activity.
D000818:Animals; D001391:Azo Compounds; D001777:Blood Coagulation / Q000187:drug effects; D002364:Caseins; D003094:Collagen; D003435:Crotalid Venoms / Q000032:analysis / Q000633:toxicity*; D004492:Edetic Acid / Q000037:antagonists & inhibitors; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005340:Fibrinogen; D006470:Hemorrhage / Q000139:chemically induced*; D006801:Humans; D006868:Hydrolysis; D066298:In Vitro Techniques; D008670:Metals / Q000494:pharmacology; D010974:Platelet Aggregation / Q000187:drug effects; D011817:Rabbits
D001391:Azo Compounds; D002364:Caseins; D003435:Crotalid Venoms; D008670:Metals; C032814:hemorrhagin components (Crotalid); C028470:Azocoll; D005340:Fibrinogen; D003094:Collagen; D004492:Edetic Acid