Purification and Characterization of Brevinase, a Heterogeneous Two-Chain Fibrinolytic Enzyme from the Venom of Korean Snake, Agkistrodon blomhoffii brevicaudus

• Published in: Biochemical and biophysical research communications Vol. 260; no. 3; pp. 665 - 670
• Main Authors: Lee, Je-Wook, Seu, Jung-Hwn, Rhee, In-Koo, Jin, Ingnyol, Kawamura, Yoshiharu, Park, Wan
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 14-07-1999
• Subjects: Agkistrodon blomhoffi brevicaudus; Agkistrodon blomhoffii brevicaudus; Amino Acid Sequence; Animals; Cations, Divalent - pharmacology; Crotalid Venoms - enzymology; Disulfides - chemistry; Disulfides - metabolism; Dithiothreitol - pharmacology; Enzyme Inhibitors - pharmacology; Enzyme Stability; Fibrinogen - metabolism; fibrinolytic serine protease; Hot Temperature; Humans; Hydrogen-Ion Concentration; Molecular Sequence Data; Molecular Weight; Sequence Analysis; Sequence Homology, Amino Acid; Serine Endopeptidases - chemistry; Serine Endopeptidases - isolation & purification; Serine Endopeptidases - metabolism; snake venom; Zinc - pharmacology; snake venom; fibrinolytic serine protease; Agkistrodon blomhoffii brevicaudus
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1006/bbrc.1999.0977

A fibrinolytic enzyme, designated as brevinase, was purified from the venom of Korean snake, Agkistrodon blomhoffii brevicaudus. Brevinase cleaved both the Aα- and Bβ-chains of fibrinogen but did not affect the γ-chain. It showed β-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°C for 3 min, showed an equal maximum activity in the pH range from 5.5 to 8.5, and was inactivated by Zn2+. 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.