Purification and characterization of a new platelet aggregation inhibitor with dissociative effect on ADP-induced platelet aggregation, from the venom of Protobothrops elegans (Sakishima-habu).
A platelet aggregation inhibitor, named snake venom platelet aggregation dissociator (SV-PAD)-1, with a dissociative reaction of ADP-induced platelet aggregation, was purified from the venom of Protobothrops elegans (Sakishima-habu) by gel-filtration employing Sephadex G-100, and ion-exchange chromatographies using DEAE-Sepharose Fast Flow, CM-Sepharose Fast Flow, and Mono S. By this procedure, about 1.5mg of purified protein was obtained from 1.0g of P. elegans venom. The purified protein showed a single protein band and the molecular weight was about 110kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under reducing conditions. The pI of purified protein showed four-bands of 7.7, 7.8, 7.95, and 8.15. This protein strongly inhibited ADP-induced platelet aggregation in rabbit platelet-rich plasma (PRP), and its IC(50) was about 58nM. It inhibited ristocetin-induced platelet aggregation in rabbit PRP (IC(50): 100nM), but hardly blocked collagen-induced platelet aggregation. This protein promptly dissociated platelet aggregation in rabbit PRP stimulated by high-concentration ADP.
Oyama
Etsuko
E
Department of Hygienic Chemistry, Meiji Pharmaceutical University, Kiyose, Tokyo, Japan. oyama@my-pharm.ac.jp
Furudate
Naomichi
N
Senuki
Kotaro
K
Takahashi
Hidenobu
H
D000244:Adenosine Diphosphate / Q000037:antagonists & inhibitors* / Q000494:pharmacology; D000818:Animals; D019921:Biotinylation; D003094:Collagen / Q000037:antagonists & inhibitors / Q000494:pharmacology; D003435:Crotalid Venoms / Q000737:chemistry / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005340:Fibrinogen / Q000737:chemistry; D066298:In Vitro Techniques; D039661:Integrin beta3 / Q000378:metabolism; D007525:Isoelectric Focusing; D008970:Molecular Weight; D010455:Peptides / Q000494:pharmacology; D010974:Platelet Aggregation / Q000187:drug effects*; D010975:Platelet Aggregation Inhibitors / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology*; D040201:Platelet Membrane Glycoprotein IIb / Q000378:metabolism; D011817:Rabbits; D012310:Ristocetin / Q000037:antagonists & inhibitors / Q000494:pharmacology; D017841:Trimeresurus / Q000502:physiology*
D003435:Crotalid Venoms; D039661:Integrin beta3; D010455:Peptides; D010975:Platelet Aggregation Inhibitors; D040201:Platelet Membrane Glycoprotein IIb; C064594:elegantin; D012310:Ristocetin; D000244:Adenosine Diphosphate; D005340:Fibrinogen; D003094:Collagen