title: Characterization of OhS1, an arginine/lysine amidase from the venom of king cobra (Ophiophagus hannah).
abstract: In this paper, we present the results of purification and characterization of an arginine/lysine amidase from the venom of Ophiophagus hannah (OhS1). It was purified by Sephadex G-75 gel filtration and ion-exchange chromatography on DEAE-Sepharose CL-6B. It is a protein of about 43,000, consisting of a single polypeptide chain. It is a minor component in the venom. The purified enzyme was capable of hydrolysing several tripeptidyl-p-nitroanilide substrates having either arginine or lysine as the C-terminal residue. We studied the kinetic parameters of OhS1 on six these chromogenic substrates. OhS1 did not clot fibrinogen. Electrophoresis of fibrinogen degraded with OhS1 revealed the disappearance of the alpha- and beta-chains and the appearance of lower mol. wt fragments. OhS1 had no hemorrhagic activity. It did not hydrolyse casein, nor did it act on blood coagulation factor X, prothrombin and plasminogen. The activity of OhS1 was completely inhibited by NPGB, PMSF, DFP, benzamidine and soybean trypsin inhibitor, suggesting it is a serine protease. Metal chelator (EDTA) had no effect on it.
authors.affiliation: Kunming Institute of Zoology, Chinese Academy of Sciences, Yunnan.
authors.forename: Y
authors.initials: Y
authors.lastname: Zhang
authors.forename: W H
authors.initials: WH
authors.lastname: Lee
authors.forename: Y L
authors.initials: YL
authors.lastname: Xiong
authors.forename: W Y
authors.initials: WY
authors.lastname: Wang
authors.forename: S W
authors.initials: SW
authors.lastname: Zu
mesh_terms: D000581:Amidohydrolases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D000595:Amino Acid Sequence; D001120:Arginine; D004546:Elapid Venoms / Q000737:chemistry*; D008239:Lysine; D008969:Molecular Sequence Data; D011480:Protease Inhibitors / Q000494:pharmacology; D012697:Serine Endopeptidases / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D004546:Elapid Venoms; D011480:Protease Inhibitors; D001120:Arginine; D012697:Serine Endopeptidases; C089175:arginine - lysine amidase, Ophiophagus hannah; D000581:Amidohydrolases; D008239:Lysine