Incorporation of the Unnatural Amino Acid p-benzoyl-L-phenylalanine (Bpa) into a G Protein-coupled Receptor in its Native Context

  • George Umanah
  • Li-Yin Huang
  • Peter G. Schultz
  • Fred Naider
  • Jeffrey M. Becker
Part of the Advances in Experimental Medicine and Biology book series (volume 611)


Ligand binding initiates a change in the conformation of G protein-coupled receptors (GPCRs) resulting in activation of the G protein-mediated signal transduction cascade [1]. We are studying a novel approach to elucidate the dynamics of GPCR structure by the co-translational introduction of unnatural amino acids (UAAs) into the receptor.

UAAs can be synthesized to contain a variety of chemical moieties for use as photoaffinity labels, fluorescent labels, or spectroscopic probes. Orthogonal tRNA/ aminoacyl-tRNA synthetase pairs evolved and expressed in the target cell have been used to incorporate UAAs into heterologously expressed protein in living cells [2]. The mutated tRNA, charged with its UAA, recognizes the amber TAG stop codon and incorporates the non-natural amino acid into the nascent polypeptide chain (See Figure below). UAAR has been widely used in the heterologous Xenopusoocyte expression system to insert UAAs into a variety of receptors and channel...


Unnatural Amino Acid Aminoacyl tRNA Synthetase Amino Acid Analog Nascent Polypeptide Chain Eukaryotic Host Cell 
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This work was supported by grants NIH GM-22086(FN) and NIH GM-22087(JMB).


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Copyright information

© Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • George Umanah
    • 1
  • Li-Yin Huang
    • 1
  • Peter G. Schultz
    • 2
  • Fred Naider
    • 3
  • Jeffrey M. Becker
    • 1
  1. 1.Department of MicrobiologyUniversity of TennesseeKnoxville
  2. 2.Department of ChemistryThe Scripps Research InstituteLa Jolla
  3. 3.Department of ChemistryCollege of Staten IslandCUNY

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