Reconstitution of G protein-coupled receptors with recombinant G protein alpha and beta gamma subunits.
The methods outlined in this article describe experiments that can probe the first steps in receptor:G protein interaction using defined, recombinant receptors and G proteins. The protocols have the advantages that the receptors are inserted properly in a cell membrane and that the investigator has complete control of the proteins reconstituted with the receptor. Specific mutations in the receptors or G proteins are studied easily and the protocols allow precise examination of the stoichiometry of the receptor:alpha:beta gamma interaction.
McIntire
William E
WE
Department of Pharmacology, University of Virginia Health System, Charlottesville, Virginia 22908, USA.
Myung
Chang-Seon
CS
MacCleery
Gavin
G
Wang
Qi
Q
Garrison
James C
JC
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002478:Cells, Cultured; D019204:GTP-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D016372:RNA, Antisense; D011956:Receptors, Cell Surface / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012680:Sensitivity and Specificity; D014162:Transfection; D014508:Urea / Q000737:chemistry
D016372:RNA, Antisense; D011956:Receptors, Cell Surface; D011994:Recombinant Proteins; D014508:Urea; D019204:GTP-Binding Proteins