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PMID: 23872075 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Constitutively active rhodopsin mutants causing night blindness are effectively phosphorylated by GRKs but differ in arrestin-1 binding.

Cellular signalling ·Vol. 25 ·No. 11 ·2013-11-00 ·Pages 2155-62

Vishnivetskiy SA, Ostermaier MK, Singhal A, Panneels V, Homan KT, Glukhova A, Sligar SG, Tesmer JJ, Schertler GF, Standfuss J, Gurevich VV

Abstract

The effects of activating mutations associated with night blindness on the stoichiometry of rhodopsin interactions with G protein-coupled receptor kinase 1 (GRK1) and arrestin-1 have not been reported. Here we show that the monomeric form of WT rhodopsin and its constitutively active mutants M257Y, G90D, and T94I, reconstituted into HDL particles are effectively phosphorylated by GRK1, as well as two more ubiquitously expressed subtypes, GRK2 and GRK5. All versions of arrestin-1 tested (WT, pre-activated, and constitutively monomeric mutants) bind to monomeric rhodopsin and show the same selectivity for different functional forms of rhodopsin as in native disc membranes. Rhodopsin phosphorylation by GRK1 and GRK2 promotes arrestin-1 binding to a comparable extent, whereas similar phosphorylation by GRK5 is less effective, suggesting that not all phosphorylation sites on rhodopsin are equivalent in promoting arrestin-1 binding. The binding of WT arrestin-1 to phospho-opsin is comparable to the binding to its preferred target, P-Rh*, suggesting that in photoreceptors arrestin-1 only dissociates after opsin regeneration with 11-cis-retinal, which converts phospho-opsin into inactive phospho-rhodopsin that has lower affinity for arrestin-1. Reduced binding of arrestin-1 to the phospho-opsin form of G90D mutant likely contributes to night blindness caused by this mutation in humans.

Keywords
Arrestin G protein-coupled receptor G protein-coupled receptor kinase GPCR GRK Monomer Nanodiscs P-Ops P-Rh P-Rh* Phosphorylation Rh Rh* WT dark phosphorylated rhodopsin dark unphosphorylated rhodopsin light-activated phosphorylated rhodopsin light-activated unphosphorylated rhodopsin phospho-opsin wild type
MeSH Terms
Animals Arrestin/genetics,metabolism Cattle Cholesterol, HDL/chemistry,metabolism G-Protein-Coupled Receptor Kinase 1/genetics,metabolism Gene Expression Regulation Isoenzymes/genetics,metabolism Mutation Night Blindness/genetics,metabolism,pathology Opsins/genetics,metabolism Phosphoproteins/genetics,metabolism Phosphorylation Protein Binding Protein Isoforms/genetics,metabolism Protein Multimerization Retinal Rod Photoreceptor Cells/metabolism,pathology Rhodopsin/genetics,metabolism Signal Transduction
Chemicals
Arrestin Cholesterol, HDL Isoenzymes Opsins Phosphoproteins Protein Isoforms Rhodopsin G-Protein-Coupled Receptor Kinase 1
Authors & Affiliations
11 authors, click to expand affiliations / ORCID
Vishnivetskiy Sergey A
Department of Pharmacology, Vanderbilt University, Nashville, TN 37232, USA.
Ostermaier Martin K
Singhal Ankita
Panneels Valerie
Homan Kristoff T
Glukhova Alisa
Sligar Stephen G
Tesmer John J G
Schertler Gebhard F X
Standfuss Joerg
Gurevich Vsevolod V
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Article Info
Journal
Cellular signalling
Abbr.
Cell Signal
ISSN
1873-3913
DOI
10.1016/j.cellsig.2013.07.009
Published
2013-11-00
Epub
2013-00-17
Pages
2155-62
Language
English
Region
England
NLM ID
8904683
PMCID
PMC3774132
Subset
IM
Grants
NHLBI NIH HHS · HL086865 · United States
NIGMS NIH HHS · R01 GM077561 · United States
NIGMS NIH HHS · R01 GM081756 · United States
NEI NIH HHS · EY011500 · United States
NHLBI NIH HHS · R01 HL086865 · United States
NIGMS NIH HHS · GM033775 · United States
NIGMS NIH HHS · GM077561 · United States
NIMH NIH HHS · MH089378 · United States
NIGMS NIH HHS · R01 GM033775 · United States
NIGMS NIH HHS · GM081756 · United States
NHLBI NIH HHS · HL071818 · United States
NEI NIH HHS · R01 EY011500 · United States
NHLBI NIH HHS · R01 HL071818 · United States
External Links
PubMed source DOI: 10.1016/j.cellsig.2013.07.009 PMC Full Text View journal details
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