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Optimizing the Genetic Incorporation of Chemical Probes into GPCRs for Photo-crosslinking Mapping and Bioorthogonal Chemistry in Live Mammalian Cells
Kapitel
- 00:04Titel
- 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
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.
