title: Purification and characterization of two high molecular mass snake venom metalloproteinases (P-III SVMPs), named SV-PAD-2 and HR-Ele-1, from the venom of Protobothrops elegans (Sakishima-habu).
abstract: We herein identified two high molecular mass metalloproteinases, named SV-PAD-2 and HR-Ele-1, in the venom of Protobothrops elegans. HR-Ele-1 appeared as a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) regard under reducing and non-reducing conditions, and the molecular mass of this protease was approximately 60 kDa under reducing conditions. On the other hand, the molecular masses of SV-PAD-2 on SDS-PAGE were 110 kDa under the non-reducing condition and 52 kDa under the reducing condition. These SVMPs exhibited fibrinogenolytic and enzymatic activities against synthetic substrates for matrix metalloproteinases (MMPs) and the insulin B-chain, and were both inhibited by EDTA. SV-PAD-2 inhibited ADP- and collagen-induced platelet aggregation, with IC50 values of 240 nM and 185 nM, respectively. HR-Ele-1 exhibited hemorrhagic activity, and its minimum hemorrhagic dose (MHD) was 0.05 μg in the guinea pig.
authors.affiliation: Department of Hygienic Chemistry, Meiji Pharmaceutical University, Japan. Electronic address: oyama@my-pharm.ac.jp.
authors.forename: Etsuko
authors.initials: E
authors.lastname: Oyama
authors.affiliation: Meiji Pharmaceutical University, Japan.
authors.forename: Hidenobu
authors.initials: H
authors.lastname: Takahashi
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000925:Anticoagulants / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D003094:Collagen / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004492:Edetic Acid / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D006168:Guinea Pigs; D006868:Hydrolysis; D020128:Inhibitory Concentration 50; D007328:Insulin / Q000378:metabolism; D045726:Metalloproteases / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010974:Platelet Aggregation / Q000187:drug effects; D017841:Trimeresurus / Q000378:metabolism*; D014757:Viper Venoms / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology
chemical_list: D000925:Anticoagulants; D007328:Insulin; D014757:Viper Venoms; D003094:Collagen; D004492:Edetic Acid; D045726:Metalloproteases
keywords: Anti-coagulant; Metalloproteinases; Snake venom