Unnatural amino acid mutagenesis of GPCRs using amber codon suppression and bioorthogonal labeling.

@article{Huber2013UnnaturalAA,
  title={Unnatural amino acid mutagenesis of GPCRs using amber codon suppression and bioorthogonal labeling.},
  author={Thomas Huber and Saranga Naganathan and He Tian and Shixin Ye and Thomas P. Sakmar},
  journal={Methods in enzymology},
  year={2013},
  volume={520},
  pages={
          281-305
        }
}
To advance dynamic, temporal, and kinetic studies of the G protein-coupled receptor (GPCR) signalosome, new approaches are required to introduce non- or minimally perturbing labels or probes into expressed receptors. We report here a series of methods that are based on unnatural amino acid mutagenesis of GPCRs using amber codon suppression technology. We show that labeling reactions at genetically introduced keto moieties (p-acetyl-L-Phe/AcF and p-benzoyl-L-Phe/BzF) are not completely… CONTINUE READING
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