Dissecting receptor-G protein specificity using G alpha chimeras.

Cabrera-Vera TM, Thomas TO, Vanhauwe J, Depree KM, Graber SG, Hamm HE
Methods Enzymol. 2002 344: 69-81

PMID: 11771421 · DOI:10.1016/s0076-6879(02)44706-4

In conclusion, by taking advantage of the overall sequence homology and structural similarity of G alpha subunits, functional chimeric G alpha subunits can be generated and used as tools for the identification of sequence-specific factors that mediate receptor: G protein specificity. The [35S]GTP gamma S binding assay and the affinity shift activity assay are two sensitive biochemical approaches that can be used to assess receptor: G protein coupling in vitro. These in vitro assays limit confounding influences from cellular proteins and allow for the strict control of receptor: G protein ratios.

MeSH Terms (19)

Animals Cattle Cell Line Cell Membrane GTP-Binding Protein alpha Subunits, Gq-G11 Guanosine 5'-O-(3-Thiotriphosphate) Heterotrimeric GTP-Binding Proteins Kinetics Protein Binding Protein Subunits Radioisotope Dilution Technique Receptors, Cell Surface Recombinant Fusion Proteins Recombinant Proteins Rod Cell Outer Segment Sensitivity and Specificity Spodoptera Sulfur Radioisotopes Transfection

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