Thrombin-like enzyme, flavovilase, with kinin-releasing activity from Trimeresurus flavoviridis (habu) venom.
A thrombin-like enzyme, flavovilase, with kinin-releasing activity was isolated, purified, and characterized from the venom of Trimeresurus flavoviridis (habu) using Sephadex G-100, DEAE-Cellulose, and CM-Cellulose column chromatographies. The final preparation was homogeneous as demonstrated by a single band on polyacrylamide gel electrophoresis, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and isoelectric focusing electrophoresis. The enzyme possesses a molecular weight of 26,500, an isoelectric point of 5.0, and consists of 247 total amino acid residues. Specific electrolytic activities of this enzyme on N-tosyl-L-arginine methyl ester (TAME) and N-benzoyl-L-arginine ethyl ester (BAEE) were determined to be 50.9 and 17.4 micromol/min/mg, respectively. The enzyme was inhibited by p-APMSF (p-amidinophenylmethanesulfonyl fluoride hydrochloride), beta-mercaptoethanol, and N-bromosuccinimide. Additionally, the enzyme was found stable to heat treatment. It was also observed that the enzyme cleaved a kininogen analog with the release of bradykinin.
Komori
Y
Y
Faculty of Pharmacy, Meijo University, Nagoya, Japan. ykomori@meijo-u.ac.jp
Tatematsu
R
R
Tanida
S
S
Nikai
T
T
D000818:Animals; D001920:Bradykinin / Q000737:chemistry; D003435:Crotalid Venoms / Q000737:chemistry* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006490:Hemostatics / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D007525:Isoelectric Focusing; D007704:Kininogens / Q000378:metabolism; D008970:Molecular Weight; D013917:Thrombin / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*
D003435:Crotalid Venoms; D006490:Hemostatics; D007704:Kininogens; C029748:Trimeresurus venoms; C441968:flavovilase; D013917:Thrombin; D001920:Bradykinin