Journal article

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

Sergey A Vishnivetskiy, Martin K Ostermaier, Ankita Singhal, Valerie Panneels, Kristoff T Homan, Alisa Glukhova, Stephen G Sligar, John JG Tesmer, Gebhard FX Schertler, Joerg Standfuss, Vsevolod V Gurevich

CELLULAR SIGNALLING | ELSEVIER SCIENCE INC | Published : 2013

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 phosphoryl..

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Awarded by NIH


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Awarded by Swiss National Science Foundation


Awarded by NATIONAL EYE INSTITUTE


Awarded by NATIONAL HEART, LUNG, AND BLOOD INSTITUTE


Awarded by NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES


Funding Acknowledgements

The authors thank Dr. T. H. Bayburt for reconstituting native bovine rhodopsin into HDL particles. This study was supported in part by NIH grants EY011500, GM077561, GM081756 (V.V.G.), HL086865, HL071818, MH089378 (JJ.G.T.), GM033775 (S.G.S.), AHA grant N014938 (K.T.H.), Swiss National Science Foundation (S.N.S.F.) grant 31003A_132815 U.S. and G.FX.S.) and 31003A_141235 (J.S.).