Effects of guanyl nucleotides and rhodopsin on ADP-ribosylation of the inhibitory GTP-binding component of adenylate cyclase by pertussis toxin.
Hormonal inhibition of adenylate cyclase is mediated by a guanyl nucleotide binding protein, Gi, which is composed of alpha, beta, and gamma subunits (Gi alpha, G beta gamma). Pertussis toxin blocks hormonal inhibition by catalyzing the ADP-ribosylation of Gi alpha. With purified Gi subunits, but without nucleotides, it was observed that toxin-catalyzed ADP-ribosylation of Gi alpha was negligible in the absence of G beta gamma; ATP, previously shown to increase ADP-ribosylation in membranes, enhanced the ADP-ribosylation of Gi alpha in the absence, more than in the presence, of G beta gamma. Prior studies (Kanaho, Y., Tsai, S.-C., Adamik, R., Hewlett, E.L., Moss, J., and Vaughan, M. (1984) J. Biol. Chem. 259, 7378-7381) had demonstrated that rhodopsin, the retinal photon receptor protein, can replace inhibitory hormone receptors, and stimulate the hydrolysis of GTP by Gi alpha in the presence of G beta gamma. Photolyzed rhodopsin, but not the inactive, dark protein, inhibited ADP-ribosylation of Gi alpha in the presence of G beta gamma. ADP-ribosylation of Gi alpha, in the presence of G beta gamma and photolyzed (but not dark) rhodopsin was increased by guanosine 5'-O-(2-thiodiphosphate) or GDP, but not by (beta, gamma-methylene)guanosine triphosphate or guanosine 5'-O-(3-thiotriphosphate). Presumably, photolyzed rhodopsin and nucleoside triphosphate analogues activate Gi, whereas with dark rhodopsin and nucleoside diphosphates Gi is in the inactive state. The latter appears to be the preferred substrate for pertussis toxin. These observations are consistent with other evidence that rhodopsin and inhibitory hormone receptors are functionally similar.
D000246:Adenosine Diphosphate Ribose / Q000378:metabolism*; D037361:Adenylate Cyclase Toxin; D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D001427:Bacterial Toxins / Q000494:pharmacology*; D003624:Darkness; D019204:GTP-Binding Proteins / Q000378:metabolism*; D006150:Guanine Nucleotides / Q000494:pharmacology*; D007700:Kinetics; D008027:Light; D009702:Nucleoside Diphosphate Sugars / Q000378:metabolism*; D037342:Pertussis Toxin; D012168:Retinal Pigments / Q000494:pharmacology*; D012243:Rhodopsin / Q000494:pharmacology*; D013329:Structure-Activity Relationship; D010566:Virulence Factors, Bordetella
D037361:Adenylate Cyclase Toxin; D001427:Bacterial Toxins; D006150:Guanine Nucleotides; D009702:Nucleoside Diphosphate Sugars; D012168:Retinal Pigments; D010566:Virulence Factors, Bordetella; D000246:Adenosine Diphosphate Ribose; D012243:Rhodopsin; D037342:Pertussis Toxin; D019204:GTP-Binding Proteins; D000262:Adenylyl Cyclases