Characterization of Neuwiedin, a new disintegrin from Bothrops neuwiedi venom gland with distinct cysteine pattern.
Disintegrins are cysteine-rich toxins containing the RGD motif exposed in a loop that binds integrins such as αIIbβ3, α5β1 and αvβ3. The flexibility of the RGD loop, controlled by the profile of the cysteine pairs and the residues flanking the RGD sequence, are key structural features for the functional activity of these molecules. Recently, our group reported a transcript in the venom gland of Bothrops neuwiedi corresponding to a new P-II SVMP precursor, BnMPIIx, in which the RGD-binding loop includes many substituted residues and unique cysteine residues at the C-terminal. In this paper, we obtained the recombinant disintegrin domain of BnMPIIx, Neuwiedin, which inhibited ADP-induced platelet aggregation, endothelial cell adhesion to fibrinogen and tube formation in Matrigel with no particular selectivity to αIIbβ3 or endothelial cell integrins. This value was also comparable to the inhibition observed with other recombinant disintegrins with conserved cysteine positions and residues in RGD loop. In this regard, Neuwiedin is an important component to understand the functional relevance of the diversity generated by accelerated evolution of venom toxins as well as to find out eventual new disintegrin-dependent targets that may be approached with disintegrins.
Laboratório de Imunopatologia, Instituto Butantan, Av. Vital Brazil, 1500, 05503-900, São Paulo, SP, Brazil.
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Lima-Dos-Santos
Laboratório de Imunopatologia, Instituto Butantan, Av. Vital Brazil, 1500, 05503-900, São Paulo, SP, Brazil.
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MS
Della-Casa
Laboratório de Imunopatologia, Instituto Butantan, Av. Vital Brazil, 1500, 05503-900, São Paulo, SP, Brazil.
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JA
Portes-Junior
Laboratório de Imunopatologia, Instituto Butantan, Av. Vital Brazil, 1500, 05503-900, São Paulo, SP, Brazil.
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Calabria
Laboratório de Imunopatologia, Instituto Butantan, Av. Vital Brazil, 1500, 05503-900, São Paulo, SP, Brazil. Electronic address: geraldo.magalhaes@butantan.gov.br.
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Magalhães
Laboratório de Imunopatologia, Instituto Butantan, Av. Vital Brazil, 1500, 05503-900, São Paulo, SP, Brazil. Electronic address: ana.moura@butantan.gov.br.
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Moura-da-Silva
D000595:Amino Acid Sequence; D000818:Animals; D017837:Bothrops; D002448:Cell Adhesion / Q000187:drug effects; D003001:Cloning, Molecular; D003435:Crotalid Venoms / Q000737:chemistry*; D003545:Cysteine / Q000737:chemistry*; D019483:Disintegrins / Q000737:chemistry*; D042783:Endothelial Cells / Q000187:drug effects; D004926:Escherichia coli / Q000235:genetics; D008969:Molecular Sequence Data; D010974:Platelet Aggregation / Q000187:drug effects; D011994:Recombinant Proteins / Q000737:chemistry; D012469:Salivary Glands / Q000378:metabolism*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D003435:Crotalid Venoms; D019483:Disintegrins; D011994:Recombinant Proteins; D003545:Cysteine
Disintegrin; Endothelial cells; Platelet; Recombinant; Snake venoms