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JoVE Journal
Chemistry

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Optimizing the Genetic Incorporation of Chemical Probes into GPCRs for Photo-crosslinking Mapping and Bioorthogonal Chemistry in Live Mammalian Cells
 

Optimizing the Genetic Incorporation of Chemical Probes into GPCRs for Photo-crosslinking Mapping and Bioorthogonal Chemistry in Live Mammalian Cells

Article DOI: 10.3791/57069-v 14:02 min April 9th, 2018
April 9th, 2018

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Summary

A facile fluorescence assay is presented to evaluate the efficiency of amino-acyl-tRNA-synthetase/tRNA pairs incorporating non-canonical amino-acids (ncAAs) into proteins expressed in mammalian cells. The application of ncAAs to study G-protein coupled receptors (GPCRs) is described, including photo-crosslinking mapping of binding sites and bioorthogonal GPCR labeling on live cells.

Tags

Keywords: GPCR Photo-crosslinking Bioorthogonal Chemistry Non-canonical Amino Acids Amber Codon Suppression Fluorescence Assay Mammalian Cells Protein-protein Interactions Ligand Binding Live Cell Imaging
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