Purification and characterization of a thrombin-like enzyme, elegaxobin, from the venom of Trimeresurus elegans (Sakishima-habu).
A thrombin-like enzyme, named elegaxobin, was purified from the venom of Trimeresurus elegans (Sakishima-habu) by gel filtration on Sephadex G-100, and ion-exchange chromatographies on Q-Sepharose Fast Flow and S-Sepharose Fast Flow. By this procedure, about 8.5 mg of purified enzyme was obtained from 1.1 g of the venom. The purified enzyme showed a single protein band in SDS-polyacrylamide electrophoresis under reducing condition and its molecular weight is 30,000. The specific activity of this enzyme toward tosyl-L-arginine methyl ester (TAME) was 490 TAME units/mg of protein. Elegaxobin clotted only rabbit fibrinogen whereas human and bovine fibrinogens were unaffected. In the fibrinogen-fibrin convertion, the enzyme released only fibrinopeptide A from rabbit fibrinogen, whereas fibrinopeptide B was not released. The N-terminal sequences (Val-Ile-Gly-Gly) of this enzyme was identical to typical sequence of serine proteinases.
Oyama
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Meiji Pharmaceutical University, 2-522-1 Noshio, Kiyose, Tokyo, Japan.
Takahashi
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H
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002265:Carboxylic Ester Hydrolases / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D003435:Crotalid Venoms / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D005340:Fibrinogen / Q000737:chemistry; D006801:Humans; D010455:Peptides / Q000737:chemistry; D011817:Rabbits; D012697:Serine Endopeptidases / Q000302:isolation & purification* / Q000378:metabolism*; D014106:Tosylarginine Methyl Ester / Q000378:metabolism; D017841:Trimeresurus / Q000378:metabolism*
D000596:Amino Acids; D003435:Crotalid Venoms; D010455:Peptides; D005340:Fibrinogen; D014106:Tosylarginine Methyl Ester; C019724:arginine esterase; D002265:Carboxylic Ester Hydrolases; D012697:Serine Endopeptidases; C408443:elegaxobin