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ISSN 0582-9879 Acta Biochim et Biophysica Sinica 2004, 36(6):425-429 CN 31-1300/Q

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Cloning and Characterization of Adinbitor, a Novel Disintegrin from the Snake Venom of Agkistrodon halys brevicaudus stejneger

Ji-Hong WANG, Yu WU, Feng REN, Li LÜ, and Bao-Chang ZHAO*

Department of Biochemistry and Molecular Biology, Dalian Medical University, Dalian 116027, China

Abstract Adinbitor was cloned from Agkistrodon halys brevicaudus stejneger and characterized as a novel disintegrin. In this study, total RNA was extracted from venom gland and used in RT-PCR to generate a cDNA which is 219 bp long. The sequence encoded a polypeptide composed of 73 amino acids, including 12 cysteines, an RGD motif, and the signature motif of disintergrin. Recombinant Adinbitor (rAdinbitor) was expressed in E. coli and purified by using the His∙Bind affinity chromatography. The IC50 for inhibiting human platelet aggregation and bFGF-induced proliferation of ECV304 cells was 6 mM and 0.89 mM respectively. Furthermore, Adinbitor significantly inhibited angiogenesis both in vivo and in vitro. Taken together, these results suggested that Adinbitor had typical functions of disintegrins.

Key words cloning; Adinbitor; disintegrin; platelet aggregation; angiogenesis

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Received: February 26, 2004 Accepted: April 7, 2004

*Corresponding author: Tel, 86-411-84720639; E-mail, [email protected]