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PMID: 20966068 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Monomeric rhodopsin is sufficient for normal rhodopsin kinase (GRK1) phosphorylation and arrestin-1 binding.

The Journal of biological chemistry ·Vol. 286 ·No. 2 ·2011-01-14 ·Pages 1420-8

Bayburt TH, Vishnivetskiy SA, McLean MA, Morizumi T, Huang CC, Tesmer JJ, Ernst OP, Sligar SG, Gurevich VV

Abstract

G-protein-coupled receptor (GPCR) oligomerization has been observed in a wide variety of experimental contexts, but the functional significance of this phenomenon at different stages of the life cycle of class A GPCRs remains to be elucidated. Rhodopsin (Rh), a prototypical class A GPCR of visual transduction, is also capable of forming dimers and higher order oligomers. The recent demonstration that Rh monomer is sufficient to activate its cognate G protein, transducin, prompted us to test whether the same monomeric state is sufficient for rhodopsin phosphorylation and arrestin-1 binding. Here we show that monomeric active rhodopsin is phosphorylated by rhodopsin kinase (GRK1) as efficiently as rhodopsin in the native disc membrane. Monomeric phosphorylated light-activated Rh (P-Rh*) in nanodiscs binds arrestin-1 essentially as well as P-Rh* in native disc membranes. We also measured the affinity of arrestin-1 for P-Rh* in nanodiscs using a fluorescence-based assay and found that arrestin-1 interacts with monomeric P-Rh* with low nanomolar affinity and 1:1 stoichiometry, as previously determined in native disc membranes. Thus, similar to transducin activation, rhodopsin phosphorylation by GRK1 and high affinity arrestin-1 binding only requires a rhodopsin monomer.

MeSH Terms
Amino Acid Sequence Animals Arrestins/genetics,metabolism Cattle Electrochemistry Fluorescence G-Protein-Coupled Receptor Kinase 1/metabolism Leucine/metabolism,pharmacology Lipids/chemistry Molecular Sequence Data Mutation Phosphorylation/physiology Protein Binding/physiology Rhodopsin/chemistry,genetics,metabolism Signal Transduction/physiology Tritium Vision, Ocular/physiology beta-Arrestins
Chemicals
Arrestins Lipids beta-Arrestins Tritium Rhodopsin G-Protein-Coupled Receptor Kinase 1 Leucine
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Bayburt Timothy H
Department of Biochemistry, University of Illinois, Urbana, Illinois 61801, USA.
Vishnivetskiy Sergey A
McLean Mark A
Morizumi Takefumi
Huang Chih-Chin
Tesmer John J G
Ernst Oliver P
Sligar Stephen G
Gurevich Vsevolod V
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Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
1083-351X
DOI
10.1074/jbc.M110.151043
Published
2011-01-14
Epub
2010-00-21
Pages
1420-8
Language
English
Region
United States
NLM ID
2985121R
PMCID
PMC3020750
Subset
IM
Grants
NIGMS NIH HHS · GM081756 · United States
NIGMS NIH HHS · R01 GM077561 · United States
NIGMS NIH HHS · R01 GM081756 · United States
NEI NIH HHS · EY011500 · United States
NIGMS NIH HHS · GM033775 · United States
NIGMS NIH HHS · GM077561 · United States
NIGMS NIH HHS · R01 GM033775 · United States
NEI NIH HHS · R01 EY011500 · United States
NHLBI NIH HHS · R01 HL071818 · United States
External Links
PubMed source DOI: 10.1074/jbc.M110.151043 PMC Full Text View journal details
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