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

Author(s)

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

Affiliation

^[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

• Document type: Journal article
• Source: JOURNAL OF PROTEOMICS; v. 113, p. 260-267, JAN 15 2015.
• Web of Science Citations: 13

Abstract

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)

• FAPESP's process: 13/07467-1 - CeTICS - Center of Toxins, Immune-Response and Cell Signaling
• Grantee: Hugo Aguirre Armelin
• Support type: Research Grants - Research, Innovation and Dissemination Centers - RIDC
• FAPESP's process: 11/08514-8 - Study of the degradome of HF3, a PIII-class snake venom metalloproteinase from Bothrops jararaca venom, on cultured fibroblasts
• Grantee: André Zelanis Palitot Pereira
• Support type: Scholarships in Brazil - Post-Doctorate
• FAPESP's process: 10/17328-0 - Comparative proteomic characterization of platelet aggregation induced by thrombin and PA-BJ, a serine proteinase from the venom of Bothrops jararaca.
• Grantee: Ana Karina de Oliveira
• Support type: Scholarships in Brazil - Doctorate
• FAPESP's process: 11/23403-8 - Study of the degradome of HF3, a PIII-class snake venom metalloproteinase from Bothrops jararaca venom, on cultured fibroblasts
• Grantee: André Zelanis Palitot Pereira
• Support type: Scholarships abroad - Research Internship - Post-doctor