Snake venom serine proteinases specificity mapping by proteomic identification of cleavage sites

Autor(es)

Zelanis, Andre ^[1] ; Huesgen, Pitter F. ^{[2, 3, 4]} ; Oliveira, Ana Karina ^{[5, 1]} ; Tashima, Alexandre K. ^[6] ; Serrano, Solange M. T. ^[1] ; Overall, Christopher M. ^{[2, 3, 7]}

Afiliação do(s) autor(es)

^[1] Inst Butantan, Lab Especial Toxinol Aplicada, Ctr Toxins Immuneresponse & Cell Signaling CeTICS, BR-05503000 Sao Paulo - Brazil

^[2] Univ British Columbia, Ctr Blood Res, Vancouver, BC V5Z 1M9 - Canada

^[3] Univ British Columbia, Dept Oral Biol & Med Sci, Vancouver, BC V5Z 1M9 - Canada

^[4] Forschungszentrum Julich, Cent Inst Engn Elect & Analyt ZEA 3, D-52425 Julich - Germany

^[5] Univ Sao Paulo, Dept Bioquim, Inst Quim, BR-05508 Sao Paulo - Brazil

^[6] Univ Fed Sao Paulo, Dept Bioquim, Sao Paulo - Brazil

^[7] Univ British Columbia, Dept Biochem & Mol Biol, Vancouver, BC V5Z 1M9 - Canada

• Tipo de documento: Artigo Científico
• Fonte: JOURNAL OF PROTEOMICS; v. 113, p. 260-267, JAN 15 2015.
• Citações Web of Science: 13

Resumo

Many snake venom toxins are serine proteases but their specific in vivo targets are mostly unknown. Various act on components of the coagulation cascade, and fibrinolytic and kallikrein-kinin systems to trigger various pathological effects observed in the envenomation. Despite showing high similarity in terms of primary structure snake venom serine proteinases (SVSPs) show exquisite specificity towards macromolecular substrates. Therefore, the characterization of their peptide bond specificity is important for understanding the active site preference associated with effective proteolysis as well as for the design of peptide substrates and inhibitors. Bothrops jararaca contains various SVSPs among which Bothrops protease A is a specific fibrinogenolytic agent and PA-BJ is a platelet-activating enzyme. In this study we used proteome derived peptide libraries in the Proteomic Identification of protease Cleavage Sites (PICS) approach to explore the peptide bond specificity of Bothrops protease A and PA-BJ in order to determine their individual peptide cleavage sequences. A total of 371 cleavage sites (208 for Bothrops protease A and 163 for PA-BJ) were detected and both proteinases displayed a clear preference for arginine at the P1 position. Moreover, the analysis of the specificity profiles of Bothrops protease A and PA-BJ revealed subtle differences in the preferences along P6-P6 `, despite a common yet unusual preference for Pro at P2. Taken together, these results map the subsite specificity of both SVSPs and shed light in the functional differences between these proteinases. Biological significance Proteolysis is key to various pathological effects observed upon envenomation by viperid snakes. The use of the Proteomic Identification of protease Cleavage Sites (PICS) approach for the easy mapping of proteinase subsite preferences at both the prime- and non-prime sides concurrently gives rise to a fresh understanding of the interaction of the snake venom senile proteinases with peptide and macromolecular substrates and indicates that their hydrolytic activity is influenced by the amino acid sequences adjacent to the scissile bond. (C) 2014 Elsevier B.V. All rights reserved. (AU)

• Processo FAPESP: 13/07467-1 - CeTICS - Centro de Toxinas, Imuno-Resposta e Sinalização Celular
• Beneficiário: Hugo Aguirre Armelin
• Modalidade de apoio: Auxílio à Pesquisa - Centros de Pesquisa, Inovação e Difusão - CEPIDs
• Processo FAPESP: 10/17328-0 - Caracterização proteômica comparativa da agregação plaquetária induzida pela trombina e pela PA-BJ, uma serinoproteinase do veneno da Bothrops jararaca.
• Beneficiário: Ana Karina de Oliveira
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 11/23403-8 - Estudo do degradoma do HF3, uma metaloproteinase hemorrágica da classe P-III do veneno da serpente Bothrops jararaca, sobre fibroblastos em cultura
• Beneficiário: André Zelanis Palitot Pereira
• Modalidade de apoio: Bolsas no Exterior - Estágio de Pesquisa - Pós-Doutorado
• Processo FAPESP: 11/08514-8 - Estudo do degradoma do HF3, uma metaloproteinase hemorrágica da classe P-III do veneno da serpente Bothrops jararaca, sobre fibroblastos em cultura
• Beneficiário: André Zelanis Palitot Pereira
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado