Investigation of rhodopsin catalyzed G-protein GTP-binding using [35S] GTP gamma S--effects of regeneration and hydroxylamine
Cook, N.J.; Pellicone, C.; Virmaux, N.
Biochemistry International 10(4): 647-653
1985
ISSN/ISBN: 0158-5231 PMID: 3927920 Document Number: 243469
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.