Purification and characterization of brevinase, a heterogeneous two-chain fibrinolytic enzyme from the venom of Korean snake, Agkistrodon blomhoffii brevicaudus.
A fibrinolytic enzyme, designated as brevinase, was purified from the venom of Korean snake, Agkistrodon blomhoffii brevicaudus. Brevinase cleaved both the Aalpha- and Bbeta-chains of fibrinogen but did not affect the gamma-chain. It showed beta-fibrinogenase activity devoid of fibrinogen clotting and caseinolytic activity. The fibrinolytic activity was completely inhibited by PMSF, DFP, Pefabloc, and DTT, indicating brevinase is a serine protease requiring disulfide bridge(s) for its activity. It kept 80% of the initial activity after heating at 100 degrees C for 3 min, showed an equal maximum activity in the pH range from 5.5 to 8.5, and was inactivated by Zn(2+). Brevinase consists of two polypeptide chains of 16.5 and 17 kDa linked by disulfide bridge(s). The N-terminal amino acid sequences of 16.5 and 17 kDa chains showed homology to the N-terminal and the internal (central region) amino acid sequences of single-chain fibrinolytic enzymes in snake venom, respectively.
Lee
J W
JW
Department of Microbiology, Kyungpook National University, Taegu, 702-701, Korea.
Seu
J H
JH
Rhee
I K
IK
Jin
I
I
Kawamura
Y
Y
Park
W
W
D000595:Amino Acid Sequence; D000818:Animals; D002413:Cations, Divalent / Q000494:pharmacology; D003435:Crotalid Venoms / Q000201:enzymology*; D004220:Disulfides / Q000737:chemistry / Q000378:metabolism; D004229:Dithiothreitol / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004795:Enzyme Stability; D005340:Fibrinogen / Q000378:metabolism*; D006358:Hot Temperature; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D008970:Molecular Weight; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid; D012697:Serine Endopeptidases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D015032:Zinc / Q000494:pharmacology
D002413:Cations, Divalent; D003435:Crotalid Venoms; D004220:Disulfides; D004791:Enzyme Inhibitors; D005340:Fibrinogen; D012697:Serine Endopeptidases; C120502:brevinase; D015032:Zinc; D004229:Dithiothreitol