NPP-BJ, a nucleotide pyrophosphatase/phosphodiesterase from Bothrops jararaca snake venom, inhibits platelet aggregation

Autor(es)

Santoro, Marcelo L. ^[1] ; Vaquero, Tais S. ^[1] ; Paes Leme, Adriana F. ^[2] ; Serrano, Solange M. T.

Afiliação do(s) autor(es)

^[1] Inst Butantan, Lab Fisiopatol, BR-05503900 Sao Paulo - Brazil

^[2] Inst Butantan, Lab Especial Toxinol Aplicada CAT CEPID, BR-05503900 Sao Paulo - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Toxicon; v. 54, n. 4, p. 499-512, SEP 15 2009.
• Citações Web of Science: 18

Resumo

Enzymes of the pyrophosphatase/phosphodiesterase family have multiple roles in extracellular nucleotide metabolism and in the regulation of nucleotide-based intercellular signaling. Snake venoms contain enzymes that hydrolyze nucleic acids and nucleotides, but their function is poorly understood. Here we describe for the first time the isolation and functional characterization of a soluble phosphodiesterase from Bothrops jararaca venom, which shows amino acid sequence similarity to mammalian nucleotide pyrophosphatase/phosphodiesterase 3 (NPP3), and inhibits ADP-induced platelet aggregation. The enzyme, named NPP-BJ, showed an apparent molecular mass of 228 kDa by size exclusion chromatography. NPP-BJ exhibited nuclease activity as well as pyrophosphatase and phosphatase activities. preferentially hydrolyzing nucleoside 5'-triphosphates over nucleoside 5'-diphosphates, but was not active upon nucleoside 5'-monophosphates. Depending on the substrate used, dithiothreitol and EDTA differently inhibited the catalytic activity of NPP-BJ. Platelet aggregation induced by ADP was also abrogated by NPP-BJ, whereas thrombin-induced platelet aggregation was only slightly attenuated. However, polyclonal antibodies raised against NPP-BJ could not abolish the lethal activity of B. jararaca venom. Altogether, these results show that NPP-BJ has a minor contribution to the lethal activity of this venom, but interferes with mechanisms of ADP-induced platelet aggregation. (C) 2009 Elsevier Ltd. All rights reserved. (AU)

• Processo FAPESP: 06/50059-8 - Proteômica aplicada ao estudo dos alvos celulares e plasmáticos do HF3, uma metaloproteinase do veneno da serpente Bothrops jararaca com atividades hemorrágica e inflamatória
• Beneficiário: Adriana Franco Paes Leme
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 04/02223-8 - Purificacao de enzimas com atividade de atpase, adpase e 5'-nucleotidase do veneno da serpente bothrops jararaca e caracterizacao de suas atividades sobre a agregacao plaquetaria.
• Beneficiário: Marcelo Larami Santoro
• Modalidade de apoio: Auxílio à Pesquisa - Regular