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Accueil du site > Animations Scientifiques > Séminaires 2010 > Shixin Ye — Application of site-directed mutagenesis with unnatural amino acids in studies of G protein-coupled receptors
Shixin Ye — Application of site-directed mutagenesis with unnatural amino acids in studies of G protein-coupled receptors
Speaker :
Shixin Ye, Laboratory of Molecular Biology and Biochemistry, The Rockefeller University, New-York
When :
Wednesday 27 October at 11am
Where :
C023 (RDC LR6 côté Centre Blaise Pascal)
Title :
Application of site-directed mutagenesis with unnatural amino acids in studies of G protein-coupled receptors
Abstract :
The introduction of unique chemical groups into proteins by means of site-directed mutagenesis with unnatural amino acids has numerous applications in protein engineering and functional studies. In this talk, I will focus on explaning the amber codon suppression technique which we have employed and optimized in rhodopsin, where the receptor gene has been co-expressed with the appropriate orthogonal pair of engineered suppressor tRNA and amino-acyl tRNA synthetase. Using this method, we have successfully introduced p-acetyl-L-phenylalanine (Acp), p-benzoyl-L-phenylalanine (Bzp) and p-azido-L-phenylalanine (Azp) into two types of G protein-coupled receptor (GPCR). The unique chemical properties of these unnatural amino acids serve as ideal an infrared probe and chemical handles for the studies of rhodopsin by FTIR and fluorescent spectroscopy. Examples of these studies will be presented.
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