title: Regulation of the methylation status of G protein-coupled receptor kinase 1 (rhodopsin kinase).
abstract: Rhodopsin kinase (GRK1) is a member of G protein-coupled receptor kinase family and a key enzyme in the quenching of photolysed rhodopsin activity and desensitisation of the rod photoreceptor neurons. Like some other rod proteins involved in phototransduction, GRK1 is posttranslationally modified at the C terminus by isoprenylation (farnesylation), endoproteolysis and α-carboxymethylation. In this study, we examined the potential mechanisms of regulation of GRK1 methylation status, which have remained unexplored so far. We found that considerable fraction of GRK1 is endogenously methylated. In isolated rod outer segments, its methylation is inhibited and demethylation stimulated by low-affinity nucleotide binding. This effect is not specific for ATP and was observed in the presence of a non-hydrolysable ATP analogue AMP-PNP, GTP and other nucleotides, and thus may involve a site distinct from the active site of the kinase. GRK1 demethylation is inhibited in the presence of Ca(2+) by recoverin. This inhibition requires recoverin myristoylation and the presence of the membranes, and may be due to changes in GRK1 availability for processing enzymes upon its redistribution to the membranes induced by recoverin/Ca(2+). We hypothesise that increased GRK1 methylation in dark-adapted rods due to elevated cytoplasmic Ca(2+) levels would further increase its association with the membranes and recoverin, providing a positive feedback to efficiently suppress spurious phosphorylation of non-activated rhodopsin molecules and thus maximise senstivity of the photoreceptor. This study provides the first evidence for dynamic regulation of GRK1 α-carboxymethylation, which might play a role in the regulation of light sensitivity and adaptation in the rod photoreceptors.
authors: Laboratoire de Biophysique Moléculaire & Cellulaire, URA CNRS N°520, Département de Biologie Moléculaire et Structurale, CEA-Grenoble, 38054 Grenoble Cedex 9, France. m.kutuzov@usa.net
authors: Mikhail A
authors: MA
authors: Kutuzov
authors: Alexandra V
authors: AV
authors: Andreeva
authors: Nelly
authors: N
authors: Bennett
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology; D000266:Adenylyl Imidodiphosphate / Q000494:pharmacology; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002417:Cattle; D051606:G-Protein-Coupled Receptor Kinase 1 / Q000378:metabolism*; D008745:Methylation / Q000187:drug effects; D010766:Phosphorylation; D051603:Recoverin / Q000494:pharmacology; D012374:Rod Cell Outer Segment / Q000378:metabolism
chemical_list: D051603:Recoverin; D000266:Adenylyl Imidodiphosphate; D000255:Adenosine Triphosphate; D051606:G-Protein-Coupled Receptor Kinase 1; D002118:Calcium