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Dissecting receptor-G protein specificity using G alpha chimeras.

Theresa M Cabrera-Vera·Tarita O Thomas·Jurgen Vanhauwe·Karyn M Depree·Stephen G Graber·Heidi E Hamm
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Abstract

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.

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AnimalsCattleCell LineCell MembraneGTP-Binding Protein alpha Subunits, Gq-G11Guanosine 5'-O-(3-Thiotriphosphate)Heterotrimeric GTP-Binding ProteinsKineticsProtein BindingProtein SubunitsRadioisotope Dilution TechniqueReceptors, Cell SurfaceRecombinant Fusion ProteinsRecombinant ProteinsRod Cell Outer SegmentSensitivity and SpecificitySpodopteraSulfur RadioisotopesTransfectionJournal Article

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Theresa M Cabrera-Vera, Tarita O Thomas, Jurgen Vanhauwe, Karyn M Depree, Stephen G Graber, Heidi E Hamm. 2002. Dissecting receptor-G protein specificity using G alpha chimeras.. https://doi.org/10.1016/s0076-6879(02)44706-4

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