title: Inhibition of rhodopsin kinase by recoverin. Further evidence for a negative feedback system in phototransduction.
abstract: Recoverin is a 23-kDa Ca(2+)-binding protein found predominantly in vertebrate photoreceptor cells. Recent electrophysiological and biochemical studies suggest that recoverin may regulate the photoresponse by inhibiting rhodopsin phosphorylation. We find in both cell homogenates and reconstituted systems that the inhibition of rhodopsin phosphorylation by recoverin occurs over a significantly higher free Ca2+ range than previously reported. Half-maximal inhibition occurs at 1.5-3 microM free Ca2+ and is cooperative with a Hill coefficient of approximately 2. Measurements of transducin activation demonstrate that this inhibition prolongs the lifetime of catalytically active rhodopsin. Ca(2+)-recoverin directly inhibits rhodopsin kinase activity, and Ca(2+)-dependent binding of recoverin to rod outer segment membranes is not required for its action. Extrapolation of the in vitro data to in vivo conditions based on simple mass action calculations places the Ca(2+)-recoverin regulation within the physiological free Ca2+ range in intact rod outer segment. The data are consistent with a model in which the fall in free Ca2+ that accompanies rod excitation exerts negative feedback by relieving inhibition of rhodopsin phosphorylation.
authors.affiliation: Laboratory of Molecular Biology, University of Wisconsin, Madison 53706, USA.
authors.forename: V A
authors.initials: VA
authors.lastname: Klenchin
authors.forename: P D
authors.initials: PD
authors.lastname: Calvert
authors.forename: M D
authors.initials: MD
authors.lastname: Bownds
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism* / Q000494:pharmacology; D002135:Calcium-Binding Proteins / Q000378:metabolism* / Q000494:pharmacology; D002417:Cattle; D004305:Dose-Response Relationship, Drug; D005136:Eye Proteins; D051606:G-Protein-Coupled Receptor Kinase 1; D051597:Hippocalcin; D008027:Light; D008074:Lipoproteins; D009419:Nerve Tissue Proteins; D010766:Phosphorylation; D011485:Protein Binding; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases; D011898:Ranidae; D051603:Recoverin; D012243:Rhodopsin / Q000378:metabolism*; D012374:Rod Cell Outer Segment / Q000378:metabolism* / Q000528:radiation effects; D015398:Signal Transduction; D013347:Subcellular Fractions / Q000378:metabolism
chemical_list: D002135:Calcium-Binding Proteins; D005136:Eye Proteins; D008074:Lipoproteins; D009419:Nerve Tissue Proteins; D047428:Protein Kinase Inhibitors; D051603:Recoverin; D051597:Hippocalcin; D012243:Rhodopsin; D011494:Protein Kinases; D051606:G-Protein-Coupled Receptor Kinase 1; D002118:Calcium