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Site-directed labeling of ß-arrestin with monobromobimane for measuring their interaction with G protein-coupled receptors.
Srivastava, Ashish; Baidya, Mithu; Dwivedi-Agnihotri, Hemlata; Shukla, Arun K.
Afiliação
  • Srivastava A; Department of Biological Sciences and Bioengineering, Indian Institute of Technology, Kanpur, India.
  • Baidya M; Department of Biological Sciences and Bioengineering, Indian Institute of Technology, Kanpur, India.
  • Dwivedi-Agnihotri H; Department of Biological Sciences and Bioengineering, Indian Institute of Technology, Kanpur, India.
  • Shukla AK; Department of Biological Sciences and Bioengineering, Indian Institute of Technology, Kanpur, India. Electronic address: arshukla@iitk.ac.in.
Methods Enzymol ; 633: 271-280, 2020.
Article em En | MEDLINE | ID: mdl-32046850
ß-arrestins (ßarrs) are multifunctional proteins that interact with activated and phosphorylated G protein-coupled receptors (GPCRs) to regulate their signaling and trafficking. Understanding the intricate details of GPCR-ßarr interaction continues to be a key research area in the field of GPCR biology. Bimane fluorescence spectroscopy has been one of the key approaches among a broad range of methods employed to study GPCR-ßarr interaction using purified and reconstituted system. Here, we present a step-by-step protocol for labeling ßarrs with monobromobimane (mBBr) in a site-directed fashion for measuring their interaction with GPCRs and the resulting conformational changes. This simple protocol can be directly applied to other protein-protein interaction modules as well for measuring interactions and conformational changes in reconstituted systems in vitro.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Compostos Bicíclicos com Pontes / Biofilmes / Reatores Biológicos / Receptores Acoplados a Proteínas G / Beta-Arrestinas Idioma: En Ano de publicação: 2020 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Compostos Bicíclicos com Pontes / Biofilmes / Reatores Biológicos / Receptores Acoplados a Proteínas G / Beta-Arrestinas Idioma: En Ano de publicação: 2020 Tipo de documento: Article