Disintegrins are among the most potent antagonists of several integrins. A cDNA encoding a novel disintegrin, bothrostatin, was cloned from a Bothrops jararaca cDNA library. The precursor of bothrostatin contains a pro, a metalloproteinase, and an RGD-disintegrin domain. The disintegrin domain expressed in Escherichia coli showed high inhibitory activity on collagen-induced platelet aggregation (IC50 of 12 nM), and thus it can be used as a useful tool for studies of integrin-ligand interaction. Furthermore, we used the comparative modeling approach to obtain a model of the 3D structure of bothrostatin. Our results suggest that bothrostatin adopts a globular, closed structure in solution. The RGD motif is exposed to the solution by the loop formed by residues 45-59 and is very close to the C-terminal domain forming a finger-like structure. The proximity of the RGD loop and the C-terminal residues, which is maintained by the Cys47-Cys66 bond, suggests that the C-terminal residues are involved in the ability of bothrostatin to interact with its ligands. (c) 2005 Elsevier Inc. All rights reserved.