title: Regulation of arrestin binding by rhodopsin phosphorylation level.
abstract: Arrestins ensure the timely termination of receptor signaling. The role of rhodopsin phosphorylation in visual arrestin binding was established more than 20 years ago, but the effects of the number of receptor-attached phosphates on this interaction remain controversial. Here we use purified rhodopsin fractions with carefully quantified content of individual phosphorylated rhodopsin species to elucidate the impact of phosphorylation level on arrestin interaction with three biologically relevant functional forms of rhodopsin: light-activated and dark phosphorhodopsin and phospho-opsin. We found that a single receptor-attached phosphate does not facilitate arrestin binding, two are necessary to induce high affinity interaction, and three phosphates fully activate arrestin. Higher phosphorylation levels do not increase the stability of arrestin complex with light-activated rhodopsin but enhance its binding to the dark phosphorhodopsin and phospho-opsin. The complex of arrestin with hyperphosphorylated light-activated rhodopsin is less sensitive to high salt and appears to release retinal faster. These data suggest that arrestin likely quenches rhodopsin signaling after the third phosphate is added by rhodopsin kinase. The complex of arrestin with heavily phosphorylated rhodopsin, which appears to form in certain disease states, has distinct characteristics that may contribute to the phenotype of these visual disorders.
authors.affiliation: Department of Pharmacology, Vanderbilt University, Nashville, Tennessee 37232, USA.
authors.forename: Sergey A
authors.initials: SA
authors.lastname: Vishnivetskiy
authors.forename: Dayanidhi
authors.initials: D
authors.lastname: Raman
authors.forename: Junhua
authors.initials: J
authors.lastname: Wei
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Kennedy
authors.forename: James B
authors.initials: JB
authors.lastname: Hurley
authors.forename: Vsevolod V
authors.initials: VV
authors.lastname: Gurevich
mesh_terms: D000818:Animals; D019393:Arrestin / Q000378:metabolism*; D001665:Binding Sites; D002417:Cattle; D051606:G-Protein-Coupled Receptor Kinase 1 / Q000378:metabolism; D010766:Phosphorylation; D012160:Retina / Q000737:chemistry / Q000378:metabolism; D012243:Rhodopsin / Q000302:isolation & purification / Q000378:metabolism*; D014785:Vision, Ocular
chemical_list: D019393:Arrestin; D012243:Rhodopsin; D051606:G-Protein-Coupled Receptor Kinase 1