Characterization of a thrombin-like enzyme from the venom of Trimeresurus jerdonii.
From the venom of Trimeresurus jerdonii, a distinct thrombin-like enzyme, called jerdonobin, was purified by DEAE A-25 ion-exchange chromatography, Sephadex G-75 gel filtration, and fast protein liquid chromatography (FPLC). SDS-PAGE analysis of this enzyme shows that it consists of a single polypeptide chain with a molecular weight of 38,000. The NH(2)-terminal amino acid sequence of jerdonobin has great homology with venom thrombin-like enzymes documented. Jerdonobin is able to hydrolyze several chromogenic substrates. The enzyme directly clots fibrinogen with an activity of 217 NIH units/mg. The fibrinopeptides released, identified by HPLC, consisted of fibrinopeptide A and a small amount of fibrinopepide B. The activities of the enzyme were inhibited by phenylmethylsulfonyl fluoride (PMSF) and p-nitrophenyl-p-guanidinobenzoate (NPGB). However, metal chelator (EDTA) had no effect on it, indicating it is venom serine protease.
Lu
Q M
QM
Department of Animal Toxinology, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223, Yunnan, China.
Jin
Y
Y
Li
D S
DS
Wang
W Y
WY
Xiong
Y L
YL
D000595:Amino Acid Sequence; D000818:Animals; D001777:Blood Coagulation / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D003435:Crotalid Venoms / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D005176:Factor XIII / Q000187:drug effects; D005337:Fibrin / Q000737:chemistry; D005340:Fibrinogen / Q000737:chemistry; D006470:Hemorrhage / Q000139:chemically induced; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010974:Platelet Aggregation / Q000187:drug effects; D012697:Serine Endopeptidases / Q000737:chemistry* / Q000302:isolation & purification; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D013056:Spectrophotometry, Ultraviolet; D013379:Substrate Specificity; D013917:Thrombin / Q000737:chemistry* / Q000302:isolation & purification; D017841:Trimeresurus
D003435:Crotalid Venoms; D015842:Serine Proteinase Inhibitors; D005337:Fibrin; D005340:Fibrinogen; D005176:Factor XIII; D012697:Serine Endopeptidases; D013917:Thrombin