Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
10880149 | Toxicon | 2011 | 9 Pages |
Abstract
Insularin (INS) was obtained from Bothrops insularis venom by reversed-phase highperformance liquid chromatography using a C18 column and characterized as a disintegrin by peptide mass fingerprint and inhibition of ADP-induced platelet aggregation. A cDNA coding for P-II a metalloproteinase/disintegrin was cloned from a cDNA library from B. insularis venom glands. The deduced protein sequence possesses 73 amino acid residues, including the N-terminal, internal peptides of native insularin, the ARGDNP-sequence and 12 cysteines in a conserved alignment. This cDNA fragment was subcloned in the pGEX-4T-1 vector and expressed in a prokaryotic expression system as a fusion protein with glutathione S-transferase (GST-INS). Both native and recombinant insularin inhibited ADP-induced platelet aggregation and endothelial cells (HUVEC) adhesion with similar activities indicating that GST-INS folded correctly and preserved the integrin-binding loop. Insularin may be a tool in studies that involve platelets and endothelial cell adhesion dependent on alphaIIbeta3 and alphavbeta3 integrins.
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Authors
Maisa Splendore Della-Casa, Inácio Junqueira-de-Azevedo, Diego Butera, PatrÃcia Bianca Clissa, Daiana S. Lopes, Solange M.T. Serrano, Daniel C. Pimenta, Geraldo S. Magalhães, Paulo Lee Ho, Ana Maria Moura-da-Silva,