A new thrombin-like enzyme, flavoviridiobin from the venom of Trimeresurus flavoviridis (habu).
Fibrinopeptide A and B releasing enzyme, flavoviridiobin, was isolated from the venom of Trimeresurus flavoviridis using Q-Sepharose, CM-Cellulose, and Sephadex G-75 column chromatographies. Homogeneity was established by the formation of a single band in polyacrylamide gel electrophoresis, isoelectric focusing, and Ouchterlony immunodiffusion. The enzyme has a molecular weight of 48,000, isoelectric point of 8.1, consists of 237 total amino acid residues, and demonstrates clotting activity. However, no tosyl-L-arginine methyl ester (TAME) hydrolytic and kinin-releasing activities were observed. This clotting enzyme was inhibited by p-amidinophenylmethanesulfonyl fluoride hydrochloride (p-APMSF), benzamidine, and beta-mercaptoethanol, suggesting that serine, acidic amino acids, and disulfide bonds are involved in the expression of the enzyme's clotting activity. This thrombin-like enzyme hydrolyzes B beta-chain of human fibrinogen at first, followed by hydrolysis A alpha-chain. The enzyme was stable over the pH range of 7-10 and was shown to be heat resistant.
Tatematsu
R
R
Department of Microbiology, Faculty of Pharmacy, Meijo University, Tenpaku-Ku, Yagotoyama 150, Nagoya 468-8503, Japan.
Komori
Y
Y
Nikai
T
T
D000818:Animals; D001777:Blood Coagulation / Q000187:drug effects; D003435:Crotalid Venoms / Q000737:chemistry / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005344:Fibrinopeptide A / Q000378:metabolism; D005345:Fibrinopeptide B / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D012697:Serine Endopeptidases / Q000302:isolation & purification*; D013696:Temperature; D017841:Trimeresurus / Q000502:physiology*
D003435:Crotalid Venoms; D005344:Fibrinopeptide A; D005345:Fibrinopeptide B; D012697:Serine Endopeptidases; C419652:flavoviridiobin