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- Title
In-Membrane Chemical Modification (IMCM) for Site-Specific Chromophore Labeling of GPCRs.
- Authors
Sušac, Lukas; O'Connor, Casey; Stevens, Raymond C.; Wüthrich, Kurt
- Abstract
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 K-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 spectroscopy experiments.
- Subjects
CHEMICAL modification of proteins; PROPERTIES of cathode rays; CRYSTAL photochemistry; DECAY-associated spectra; FLUORESCENCE aided caries excavation; OPIOID peptides
- Publication
Angewandte Chemie, 2015, Vol 127, Issue 50, p15461
- ISSN
0044-8249
- Publication type
Article
- DOI
10.1002/ange.201508506