M opsin phosphorylation in intact mammalian retinas.
The deactivation of visual pigments involved in phototransduction is critical for recovering sensitivity after exposure to light in rods and cones of the vertebrate retina. In rods, phosphorylation of rhodopsin by rhodopsin kinase (GRK1) and the subsequent binding of visual arrestin completely terminates phototransduction. Although signal termination in cones is predicted to occur via a similar mechanism as in rods, there may be differences due to the expression of related but distinct gene products. While rods only express GRK1, cones in some species express only GRK1 or GRK7 and others express both GRKs. In the mouse, cone opsin is phosphorylated by GRK1, but this has not been demonstrated in mammals that express GRK7 in cones. We compared cone opsin phosphorylation in intact retinas from the 13-lined ground squirrel (GS) and pig, cone- and rod-dominant mammals, respectively, which both express GRK7. M opsin phosphorylation increased during continuous exposure to light, then declined between 3 and 6 min. In contrast, rhodopsin phosphorylation continued to increase during this time period. In GS retina homogenates, anti-GS GRK7 antibody blocked M opsin phosphorylation by 73%. In pig retina homogenates, only 20% inhibition was observed, possibly due to phosphorylation by GRK1 released from rods during homogenization. Our results suggest that GRK7 phosphorylates M opsin in both of these mammals. Using an in vitro GTPgammaS binding assay, we also found that the ability of recombinant M opsin to activate G(t) was greatly reduced by phosphorylation. Therefore, phosphorylation may participate directly in the termination of phototransduction in cones by decreasing the activity of M opsin.
Liu
Peng
P
Department of Cell and Developmental Biology, The University of North Carolina at Chapel Hill, North Carolina 27599-7090, USA.
Osawa
Shoji
S
Weiss
Ellen R
ER
D000222:Adaptation, Physiological; D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D015153:Blotting, Western / Q000379:methods; D002460:Cell Line; D003001:Cloning, Molecular / Q000379:methods; D054768:G-Protein-Coupled Receptor Kinases; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000378:metabolism; D006801:Humans; D047468:Immunoprecipitation / Q000379:methods; D008027:Light; D010759:Phosphorus Isotopes / Q000494:pharmacology; D010766:Phosphorylation / Q000528:radiation effects; D017346:Protein Serine-Threonine Kinases / Q000276:immunology / Q000378:metabolism; D012160:Retina / Q000378:metabolism* / Q000528:radiation effects; D012243:Rhodopsin / Q000378:metabolism; D012374:Rod Cell Outer Segment / Q000378:metabolism; D017299:Rod Opsins / Q000276:immunology / Q000378:metabolism*; D012589:Sciuridae; D013552:Swine; D013997:Time Factors; D014162:Transfection / Q000379:methods
D000906:Antibodies; D010759:Phosphorus Isotopes; D017299:Rod Opsins; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D000255:Adenosine Triphosphate; D012243:Rhodopsin; D017346:Protein Serine-Threonine Kinases; C111571:GRK7 protein, human; D054768:G-Protein-Coupled Receptor Kinases