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