In-Membrane Chemical Modification (IMCM) for Site-Specific Chromophore Labeling of GPCRs
2015
We present in-membrane
chemical modification(IMCM) for obtaining selective
chromophorelabeling 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
chromophoresfor NMR and
fluorescence spectroscopyin the human
κ-opioid receptor(KOR) and the human A2A
adenosine receptor(A2AAR). 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
spectroscopyexperiments. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
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