Purification and characterization of piscivorase I and II, the fibrinolytic enzymes from eastern cottonmouth moccasin venom (Agkistrodon piscivorus piscivorus).
Fibrinolytic enzymes, piscivorase I and II, were isolated from Agkistrodon piscivorus piscivorus (eastern cottonmouth moccasin) venom using gel filtration on Bio-Gel P-100 and ion-exchange chromatography on CM-Sepharose CL-6B. The mol. wts of these proteases, piscivorase I and II, are 23,400 and 29,000 and isoelectric points are 6.6 and 8.5, respectively. These fibrinolytic enzymes were homogeneous by SDS-polyacrylamide gel electrophoresis. Piscivorase I readily cleaved the A alpha- and B beta-chain of fibrinogen, but piscivorase II cleaved readily the A alpha-chain and more slowly the B beta-chain. These fibrinolytic enzymes were activated by Ca2+, Mg2+ and Ba2+, but inhibited by Zn2+, Cu2+ and Mn2+. Both fibrinolytic enzymes were also inhibited by cysteine, beta-mercaptoethanol, and by metal chelators such as EDTA and EGTA, but not by benzamidine, phenylmethanesulfonyl fluoride (PMSF), soybean trypsin inhibitor and aprotinin. These fibrinolytic enzymes did not act like thrombin, plasmin and kallikrein, using specific chromogenic substrates. Neither fibrinolytic enzyme induced platelet aggregation, and piscivorase I showed low haemorrhagic activity at dosages of 55 micrograms.
Hahn
B S
BS
Natural Products Research Institute, Seoul National University, South Korea.
Chang
I M
IM
Kim
Y S
YS
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002863:Chromogenic Compounds / Q000737:chemistry; D003435:Crotalid Venoms / Q000737:chemistry / Q000201:enzymology* / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005340:Fibrinogen / Q000187:drug effects; D005341:Fibrinolysin / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D005342:Fibrinolysis / Q000187:drug effects; D006470:Hemorrhage / Q000139:chemically induced; D006863:Hydrogen-Ion Concentration; D008670:Metals / Q000494:pharmacology; D010974:Platelet Aggregation / Q000187:drug effects; D013696:Temperature
C031442:Agkistrodon venoms; D000596:Amino Acids; D002863:Chromogenic Compounds; D003435:Crotalid Venoms; D004791:Enzyme Inhibitors; D008670:Metals; D005340:Fibrinogen; D005341:Fibrinolysin