Busca avançada
Ano de início
Entree
(Referência obtida automaticamente do Web of Science, por meio da informação sobre o financiamento pela FAPESP e o número do processo correspondente, incluída na publicação pelos autores.)

Peptidomics of Three Bothrops Snake Venoms: Insights Into the Molecular Diversification of Proteomes and Peptidomes

Texto completo
Autor(es):
Tashima, Alexandre K. [1, 2] ; Zelanis, Andre [1] ; Kitano, Eduardo S. [1] ; Ianzer, Danielle [1] ; Melo, Robson L. [1] ; Rioli, Vanessa [1] ; Sant'anna, Savio S. [3] ; Schenberg, Ana C. G. [4] ; Camargo, Antonio C. M. [1] ; Serrano, Solange M. T. [1]
Número total de Autores: 10
Afiliação do(s) autor(es):
[1] Inst Butantan, Lab Especial Toxinol Aplicada, CAT Cepid, BR-05503000 Sao Paulo - Brazil
[2] Univ Fed Sao Paulo, Dept Ciencias Exatas & Terra, Diadema - Brazil
[3] Inst Butantan, Lab Herpetol, BR-05503000 Sao Paulo - Brazil
[4] Univ Sao Paulo, Inst Ciencias Biomed, BR-05508 Sao Paulo - Brazil
Número total de Afiliações: 4
Tipo de documento: Artigo Científico
Fonte: MOLECULAR & CELLULAR PROTEOMICS; v. 11, n. 11, p. 1245-1262, NOV 2012.
Citações Web of Science: 48
Resumo

Snake venom proteomes/peptidomes are highly complex and maintenance of their integrity within the gland lumen is crucial for the expression of toxin activities. There has been considerable progress in the field of venom proteomics, however, peptidomics does not progress as fast, because of the lack of comprehensive venom sequence databases for analysis of MS data. Therefore, in many cases venom peptides have to be sequenced manually by MS/MS analysis or Edman degradation. This is critical for rare snake species, as is the case of Bothrops cotiara (BC) and B. fonsecai (BF), which are regarded as near threatened with extinction. In this study we conducted a comprehensive analysis of the venom peptidomes of BC, BF, and B. jararaca (BJ) using a combination of solid-phase extraction and reversed-phase HPLC to fractionate the peptides, followed by nano-liquid chromatography-tandem MS (LC-MS/MS) or direct infusion electrospray ionization-(ESI)-MS/MS or MALDI-MS/MS analyses. We detected marked differences in the venom peptidomes and identified peptides ranging from 7 to 39 residues in length by de novo sequencing. Forty-four unique sequences were manually identified, out of which 30 are new peptides, including 17 bradykinin-potentiating peptides, three poly-histidine-poly-glycine peptides and interestingly, 10 L-amino acid oxidase fragments. Some of the new bradykinin-potentiating peptides display significant bradykinin potentiating activity. Automated database search revealed fragments from several toxins in the peptidomes, mainly from L-amino acid oxidase, and allowed the determination of the peptide bond specificity of proteinases and amino acid occurrences for the P4-P4' sites. We also demonstrate that the venom lyophilization/resolubilization process greatly increases the complexity of the peptidome because of the imbalance caused to the venom proteome and the consequent activity of proteinases on venom components. The use of proteinase inhibitors clearly showed different outcomes in the peptidome characterization and suggested that degradomic-peptidomic analysis of snake venoms is highly sensitive to the conditions of sampling procedures. Molecular \& Cellular Proteomics 11: 10.1074/mcp.M112.019331, 1245-1262, 2012. (AU)

Processo FAPESP: 98/14307-9 - Center for Applied Toxinology
Beneficiário:Hugo Aguirre Armelin
Linha de fomento: Auxílio à Pesquisa - Centros de Pesquisa, Inovação e Difusão - CEPIDs
Processo FAPESP: 07/54626-7 - Proteômica e transcriptômica aplicadas ao estudo das variações na composição do veneno de serpentes do gênero Bothrops relacionadas à idade e ao sexo
Beneficiário:Solange Maria de Toledo Serrano
Linha de fomento: Auxílio à Pesquisa - Regular
Processo FAPESP: 11/10468-4 - HUPO 2011 - 10th world Congress
Beneficiário:Alexandre Keiji Tashima
Linha de fomento: Auxílio à Pesquisa - Reunião - Exterior
Processo FAPESP: 09/15932-0 - III congresso da BrMass - 2009
Beneficiário:Alexandre Keiji Tashima
Linha de fomento: Auxílio à Pesquisa - Reunião - Brasil
Processo FAPESP: 04/14846-0 - Rede de proteoma do estado de São Paulo
Beneficiário:Fabio Cesar Gozzo
Linha de fomento: Auxílio à Pesquisa - Programa GENOMA