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Molecular characterization of an acidic phospholipase A2 from Bothrops pirajai snake venom: synthetic C-terminal peptide identifies its antiplatelet region

  • Molecular Toxicology
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Abstract

This paper describes a biochemical and pharmacological characterization of BpirPLA2-I, the first acidic Asp49-PLA2 isolated from Bothrops pirajai. BpirPLA2-I caused hypotension in vivo, presented phospholipolytic activity upon artificial substrates and inhibitory effects on platelet aggregation in vitro. Moreover, a synthetic peptide of BpirPLA2-I, comprising residues of the C-terminal region, reproduced the antiplatelet activity of the intact protein. A cDNA fragment of 366 bp encompassing the mature form of BpirPLA2-I was cloned by reverse transcriptase-PCR of B. pirajai venom gland total RNA. A Bayesian phylogenetic analysis indicated that BpirPLA2-I forms a clade with other acid Asp49-PLA2 enzymes from the Bothrops genus, which are characterized by the high catalytic activity associated with anticoagulant or hypotensive activity or both. Comparison of the electrostatic potential (EP) on the molecular surfaces calculated from a BpirPLA2-I homology model and from the crystallographic models of a group of close homologues revealed that the greatest number of charge inversions occurred on the face opposite to the active site entrance, particularly in the Ca2+ ion binding loop. This observation suggests a possible relationship between the basic or acid character of PLA2 enzymes and the functionality of the Ca2+ ion binding loop.

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Acknowledgments

This work was supported by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) and Instituto Nacional de Ciência e Tecnologia em Toxinas (INCT-Tox), Brazil. We are grateful to Prof. Dr. J. R. Giglio (FMRP-USP) for their manuscript revision; Prof. Dr. A. Nomizo (FCFRP-USP, Brazil), MSc J. Fernández and Prof. Dr. B. Lomonte (ICP, UCR, Costa Rica) for their collaboration in the cytotoxic and hypotension assays. Thank to E. A. Bastos (TT-FAPESP) for their helpful technical collaboration.

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Authors and Affiliations

  1. Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, FCFRP-USP, Ribeirão Preto, SP, Brazil

    Sabrina S. Teixeira, Lucas B. Silveira, Franco M. N. da Silva, Daniela P. Marchi-Salvador, Luiz Fernando M. Izidoro, Suely V. Sampaio & Andreimar M. Soares

  2. Departamento de Biologia Molecular, Centro de Ciências Exatas e da Natureza, DBM-UFPB, João Pessoa, PB, Brazil

    Daniela P. Marchi-Salvador

  3. Laboratório de Bioquímica de Proteínas e Peptídeos, Instituto Oswaldo Cruz, IOC-FIOCRUZ, Rio de Janeiro, RJ, Brazil

    Floriano P. Silva Jr

  4. Faculdade de Ciências Integradas do Pontal, UFU, Ituiutaba, MG, Brazil

    Luiz Fernando M. Izidoro

  5. Departamento de Biologia Celular e Molecular (GCM), Instituto de Biologia, UFF, Niterói, RJ, Brazil

    André L. Fuly

  6. Departamento de Biofísica, UNIFESP, São Paulo, SP, Brazil

    Maria A. Juliano

  7. Laboratório Nacional de Biociências, Centro Nacional de Pesquisas em Energia e Materiais, Campinas, SP, Brazil

    Camila R. dos Santos & Mário T. Murakami

  8. Universidade Federal de São João Del Rei, UFSJ Campus, Divinópolis, MG, Brazil

    Saulo L. da Silva

  9. Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de Sao Paulo—USP, Avenida do Café, s/n°, 14040-903, Ribeirão Preto, SP, Brazil

    Andreimar M. Soares

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  1. Sabrina S. Teixeira
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Correspondence to Andreimar M. Soares.

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Teixeira, S.S., Silveira, L.B., da Silva, F.M.N. et al. Molecular characterization of an acidic phospholipase A2 from Bothrops pirajai snake venom: synthetic C-terminal peptide identifies its antiplatelet region. Arch Toxicol 85, 1219–1233 (2011). https://doi.org/10.1007/s00204-011-0665-6

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