title: Dissecting receptor-G protein specificity using G alpha chimeras.
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.
authors.affiliation: Department of Molecular Pharmacology and Biochemistry, Institute for Neuroscience, Northwestern University, Chicago, Illinois 60611, USA.
authors.forename: Theresa M
authors.initials: TM
authors.lastname: Cabrera-Vera
authors.forename: Tarita O
authors.initials: TO
authors.lastname: Thomas
authors.forename: Jurgen
authors.initials: J
authors.lastname: Vanhauwe
authors.forename: Karyn M
authors.initials: KM
authors.lastname: Depree
authors.forename: Stephen G
authors.initials: SG
authors.lastname: Graber
authors.forename: Heidi E
authors.initials: HE
authors.lastname: Hamm
mesh_terms: D000818:Animals; D002417:Cattle; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D043802:GTP-Binding Protein alpha Subunits, Gq-G11; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000378:metabolism; D020962:Heterotrimeric GTP-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D011485:Protein Binding; D021122:Protein Subunits; D011865:Radioisotope Dilution Technique; D011956:Receptors, Cell Surface / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D012374:Rod Cell Outer Segment / Q000378:metabolism; D012680:Sensitivity and Specificity; D018411:Spodoptera; D013462:Sulfur Radioisotopes; D014162:Transfection
chemical_list: D021122:Protein Subunits; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D013462:Sulfur Radioisotopes; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D043802:GTP-Binding Protein alpha Subunits, Gq-G11; D020962:Heterotrimeric GTP-Binding Proteins