Dissecting receptor-G protein specificity using G alpha chimeras.
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.
Cabrera-Vera
Theresa M
TM
Department of Molecular Pharmacology and Biochemistry, Institute for Neuroscience, Northwestern University, Chicago, Illinois 60611, USA.
Thomas
Tarita O
TO
Vanhauwe
Jurgen
J
Depree
Karyn M
KM
Graber
Stephen G
SG
Hamm
Heidi E
HE
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
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