A new short chain RGD-containing disintegrin, accutin, inhibits the common pathway of human platelet aggregation

• Published in: Biochimica et biophysica acta Vol. 1425; no. 3; pp. 493 - 504
• Main Authors: Yeh, Chia-Hsin, Peng, Hui-Chin, Yih, Jyi-Bang, Huang, Tur-Fu
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 27.11.1998
• Subjects: Accutin; Amino Acid Sequence; Amino Acids - analysis; Binding, Competitive; Blood Platelets - metabolism; Chromatography, Gel; Crotalid Venoms - chemistry; Crotalid Venoms - isolation & purification; Crotalid Venoms - metabolism; Disintegrin; Disintegrins - chemistry; Disintegrins - isolation & purification; Disintegrins - metabolism; Endothelium, Vascular - metabolism; Fluorescein-5-isothiocyanate; Glycoprotein IIb/IIIa; Humans; Molecular Sequence Data; Oligopeptides - analysis; Peptides - metabolism; Platelet aggregation inhibitor; Platelet Aggregation Inhibitors - isolation & purification; Platelet Glycoprotein GPIIb-IIIa Complex - antagonists & inhibitors; Receptors, Vitronectin - antagonists & inhibitors; RGD-containing peptide; Snake venom protein; Platelet aggregation inhibitor; Snake venom protein; RGD-containing peptide; Disintegrin; Accutin; Glycoprotein IIb/IIIa
• ISSN: 0304-4165; 0006-3002; 1872-8006
• DOI: 10.1016/S0304-4165(98)00104-4

A new short-chain disintegrin, accutin, was purified from the Formosan Agkistrodon acutus venom by using of gel filtration, ion exchanger and reverse phase HPLC. The homogeneous protein is a 47-residue polypeptide with a molecular mass of 5241 Da containing an Arg-Gly-Asp sequence and seven cysteinyl residues at positions highly homologous to other disintegrins. Accutin dose-dependently inhibited human platelet aggregation stimulated by ADP, collagen, thrombin or the thromboxane analogue U46619 in platelet suspension with IC 50 values of 66–267 nM. It was also active in inhibiting platelet aggregation of platelet-rich plasma. However, accutin apparently did not affect the shape change caused by these agonists. Accutin also inhibited fibrinogen-induced aggregation of human elastase-treated platelets in a dose-dependent manner. Furthermore, accutin dose-dependently inhibited the binding reaction of fluorescein isothiocyanate (FITC)-conjugated arietin, a member of the disintegrin family, to human platelets. In addition, the binding of FITC-conjugated accutin to platelets was almost completely blocked by a monoclonal antibody, 7E3, raised against the platelet glycoprotein IIb/IIIa complex. On the other hand, accutin as well as other disintegrins, rhodostomin and arietin, exhibited an inhibitory effect on 7E3 binding toward platelets and endothelial cells in a dose-dependent manner. It is concluded that accutin, a new platelet aggregation inhibitor belonging to the short-chain disintegrin family, acts specifically on a binding epitope of GPIIb/IIIa overlapping with that of 7E3, leading to the blockade of fibrinogen binding to its receptor.