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

14:02 min

April 09, 2018

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Chapters

  • 00:04Title
  • 01:11Fluorescence-based Screening of Incorporation Efficiencies
  • 04:22Genetic Incorporation of ncAAs into GPCRs
  • 08:21Ultrafast Bioorthogonal Labeling of GPCRs on Live Mammalian Cells
  • 11:11Results: Incorporation of Chemical Probes into GPCRs in Mammalian Cells for Cross-linking and Imaging Studies
  • 12:52Conclusion

Summary

Automatic Translation

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.

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