Abstract:
:β-arrestins (βarrs) 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-βarr 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-βarr interaction using purified and reconstituted system. Here, we present a step-by-step protocol for labeling βarrs 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.
journal_name
Methods Enzymoljournal_title
Methods in enzymologyauthors
Srivastava A,Baidya M,Dwivedi-Agnihotri H,Shukla AKdoi
10.1016/bs.mie.2019.11.009subject
Has Abstractpub_date
2020-01-01 00:00:00pages
271-280eissn
0076-6879issn
1557-7988pii
S0076-6879(19)30466-5journal_volume
633pub_type
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