Investigation of rhodopsin catalyzed G-protein GTP-binding using [35S] GTP gamma S--effects of regeneration and hydroxylamine.
A simple reconstitution system for studying rhodopsin-catalyzed G-protein GTP-binding is described. Purified rhodopsin is recombined with a G-protein containing extract in the presence of [35S]GTP gamma S and after incubation the reaction is stopped by rapid cooling and filtration. The system has been used to study the effects of 11-cis-retinal and hydroxylamine on R*-catalyzed G-protein GTP-binding. Since both these compounds greatly reduced binding, our results provide further evidence that it is the rhodopsin intermediate metarhodopsin II which interacts with G-protein.
D000818:Animals; D002417:Cattle; D019204:GTP-Binding Proteins / Q000378:metabolism*; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D006160:Guanosine Triphosphate / Q000031:analogs & derivatives / Q000378:metabolism*; D019811:Hydroxylamine; D006898:Hydroxylamines / Q000494:pharmacology*; D008027:Light; D010786:Photoreceptor Cells / Q000378:metabolism*; D012168:Retinal Pigments / Q000378:metabolism*; D012172:Retinaldehyde / Q000494:pharmacology; D012243:Rhodopsin / Q000378:metabolism*; D012374:Rod Cell Outer Segment / Q000378:metabolism*; D013873:Thionucleotides; D014785:Vision, Ocular
D006898:Hydroxylamines; D012168:Retinal Pigments; D013873:Thionucleotides; D019811:Hydroxylamine; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D006160:Guanosine Triphosphate; D012243:Rhodopsin; D019204:GTP-Binding Proteins; D012172:Retinaldehyde