Phosphorylation of rhodopsin and quenching of cyclic GMP phosphodiesterase activation by ATP at weak bleaches.
Light and GTP-dependent cyclic GMP phosphodiesterase activation of rod disk membranes is rapidly quenched by ATP. Maximum speed of this effect occurs only with the weakest bleaches. Though it has been proposed that ATP mediates its effect through rapid phosphorylation of bleached rhodopsin, previous workers have found phosphorylation kinetics too slow by more than an order of magnitude to be causal in quenching of cyclic GMP phosphodiesterase activation. In this report, we use preparations retaining more endogenous rhodopsin kinase, higher specific activity ATP, and cyclic GMP phosphodiesterase quenching conditions to show that ATP-dependent multiple phosphorylation of rhodopsin at very weak bleaches (10(-5)) is complete in less than 2 s, easily compatible with cyclic GMP phosphodiesterase quench times of 4 s measured under identical conditions. Thus, it seems likely that previous efforts to achieve high 32P counts by using large bleaches have produced conditions of substrate saturation where much longer times to completion are caused by a very large ratio of substrate to enzyme velocity. Such conditions are not appropriately compared to those that support rapid quenching. We conclude that the speed of rhodopsin phosphorylation is, in fact, adequate to explain ATP quenching of cyclic GMP phosphodiesterase activation.
D015106:3',5'-Cyclic-GMP Phosphodiesterases / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000494:pharmacology*; D000818:Animals; D002417:Cattle; D002462:Cell Membrane / Q000201:enzymology; D005136:Eye Proteins; D051606:G-Protein-Coupled Receptor Kinase 1; D006160:Guanosine Triphosphate / Q000494:pharmacology; D007700:Kinetics; D008027:Light; D010766:Phosphorylation; D010786:Photoreceptor Cells / Q000201:enzymology*; D011494:Protein Kinases / Q000378:metabolism*; D012168:Retinal Pigments / Q000378:metabolism*; D012243:Rhodopsin / Q000378:metabolism*
D005136:Eye Proteins; D012168:Retinal Pigments; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D012243:Rhodopsin; D011494:Protein Kinases; D051606:G-Protein-Coupled Receptor Kinase 1; D015106:3',5'-Cyclic-GMP Phosphodiesterases