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Two-Dimensional Blue Native/SDS Polyacrylamide Gel Electrophoresis for Analysis of Brazilian Bothrops Snake Venoms.
Ferreira de Oliveira, Natacha; Sachetto, Ana Teresa Azevedo; Santoro, Marcelo Larami.
Afiliación
  • Ferreira de Oliveira N; Laboratório de Fisiopatologia, Instituto Butantan, São Paulo 05503-900, SP, Brazil.
  • Sachetto ATA; Escola Superior do Instituto Butantan (ESIB), Instituto Butantan, São Paulo 05503-900, SP, Brazil.
  • Santoro ML; Laboratório de Fisiopatologia, Instituto Butantan, São Paulo 05503-900, SP, Brazil.
Toxins (Basel) ; 14(10)2022 Sep 23.
Article en En | MEDLINE | ID: mdl-36287928
Viperidae snakes are the most important agents of snakebites in Brazil. The protein composition of snake venoms has been frequently analyzed by means of electrophoretic techniques, but the interaction of proteins in venoms has barely been addressed. An electrophoretic technique that has gained prominence to study this type of interaction is blue native polyacrylamide gel electrophoresis (BN-PAGE), which allows for the high-resolution separation of proteins in their native form. These protein complexes can be further discriminated by a second-dimension gel electrophoresis (SDS-PAGE) from lanes cut from BN-PAGE. Once there is no study on the use of bidimensional BN/SDS-PAGE with snake venoms, this study initially standardized the BN/SDS-PAGE technique in order to evaluate protein interactions in Bothrops atrox, Bothrops erythromelas, and Bothrops jararaca snake venoms. Results of BN/SDS-PAGE showed that native protein complexes were present, and that snake venom metalloproteinases and venom serine proteinases maintained their enzymatic activity after BN/SDS-PAGE. C-type lectin-like proteins were identified by Western blotting. Therefore, bidimensional BN/SDS-PAGE proved to be an easy, practical, and efficient method for separating functional venom proteins according to their assemblage in complexes, as well as to analyze their biological activities in further details.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Bothrops / Venenos de Crotálidos Límite: Animals País/Región como asunto: America do sul / Brasil Idioma: En Revista: Toxins (Basel) Año: 2022 Tipo del documento: Article País de afiliación: Brasil Pais de publicación: Suiza

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Bothrops / Venenos de Crotálidos Límite: Animals País/Región como asunto: America do sul / Brasil Idioma: En Revista: Toxins (Basel) Año: 2022 Tipo del documento: Article País de afiliación: Brasil Pais de publicación: Suiza