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In‐Membrane Chemical Modification (IMCM) for Site‐Specific Chromophore Labeling of GPCRs
Author(s) -
Sušac Lukas,
O'Connor Casey,
Stevens Raymond C.,
Wüthrich Kurt
Publication year - 2015
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201508506
Subject(s) - g protein coupled receptor , chromophore , chemistry , membrane , fluorescence spectroscopy , biophysics , intracellular , receptor , fluorescence , combinatorial chemistry , biochemistry , photochemistry , biology , physics , quantum mechanics
We present in‐membrane chemical modification (IMCM) for obtaining selective chromophore labeling of intracellular surface cysteines in G‐protein‐coupled receptors (GPCRs) with minimal mutagenesis. This method takes advantage of the natural protection of most cysteines by the membrane environment. Practical use of IMCM is illustrated with the site‐specific introduction of chromophores for NMR and fluorescence spectroscopy in the human κ‐opioid receptor (KOR) and the human A 2A adenosine receptor (A 2A AR). IMCM is applicable to a wide range of in vitro studies of GPCRs, including single‐molecule spectroscopy, and is a promising platform for in‐cell spectroscopy experiments.