Navegando por Palavras-chave "Proteome derived peptide library"

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    Proteome-derived peptide library for the elucidation of the cleavage specificity of HF3, a snake venom metalloproteinase
    (Springer Wien, 2016) Bertholim, Luciana; Zelanis, Andre [UNIFESP]; Oliveira, Ana K.; Serrano, Solange M. T.
    The Proteomic Identification of Cleavage Sites (PICS) approach was employed for profiling the substrate specificity of HF3, a hemorrhagic snake venom metalloproteinase (SVMP) from Bothrops jararaca. A tryptic peptide library from human plasma was subject to HF3 cleavage and amino acid occurrence for P6 to P6' sites was mapped. 71 cleavage sites were detected and revealed a clear preference for leucine at P1' position, followed by hydrophobic residues in P2'. PICS confirmed existing data on prime site specificity of SVMPs.
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    Snake venom serine proteinases specificity mapping by proteomic identification of cleavage sites
    (Elsevier B.V., 2015-01-15) Zelanis, Andre; Huesgen, Pitter F.; Oliveira, Ana Karina; Tashima, Alexandre K. [UNIFESP]; Serrano, Solange M. T.; Overall, Christopher M.; Inst Butantan; Univ British Columbia; Forschungszentrum Julich; Universidade de São Paulo (USP); Universidade Federal de São Paulo (UNIFESP)
    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 significanceProteolysis 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.