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Genetically-encoded Molecular Probes to Study G Protein-coupled Receptors
JoVE Journal
Biology
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JoVE Journal Biology
Genetically-encoded Molecular Probes to Study G Protein-coupled Receptors
DOI:

16:16 min

September 13, 2013

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Chapters

  • 00:05Title
  • 02:23Site-specific Genetic Incorporation of Unnatural Amino Acids into GPCRs
  • 03:15Mapping a Ligand Binding Site Using Targeted Photocrosslinking
  • 07:12Targeted Epitope Tagging and Cell Surface Enzyme-linked Immunosorbent Assay (ELISA)
  • 10:23Bioorthogonal Fluorescent Labeling of a GPCR
  • 12:10Results: Versatile Applications of Site-specific azF Incorporation into GPCRs

Summary

Automatic Translation

We genetically-encode the unnatural amino acid, p-azido-L-phenylalanine at various targeted positions in GPCRs and show the versatility of the azido group in different applications. These include a targeted photocrosslinking technology to identify residues in the ligand-binding pocket of a GPCR, and site-specific bioorthogonal modification of GPCRs with a peptide-epitope tag or fluorescent probe.

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