Rhodopsin Arginine-135 Mutants Are Phosphorylated by Rhodopsin Kinase and Bind Arrestin in the Absence of 11-cis-Retinal

Arginine-135, located at the border between the third transmembrane domain and the second cytoplasmic loop of rhodopsin, is one of the most highly conserved amino acids in the family of G protein-coupled receptors. The effect of mutation at Arg-135 on the ability of rhodopsin to undergo desensitizat...

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Bibliographic Details
Published in:Biochemistry (Easton) 1998-04, Vol.37 (14), p.4869-4874
Main Authors: Shi, Wen, Sports, Charlene D, Raman, Dayanidhi, Shirakawa, Satoko, Osawa, Shoji, Weiss, Ellen R
Format: Article
Language:English
Subjects:
Animals
Arginine - genetics
Arginine - metabolism
Arrestin - metabolism
Cattle
Cell Line
Eye Proteins
G-Protein-Coupled Receptor Kinase 1
GTP-Binding Proteins - metabolism
Humans
Mutagenesis, Site-Directed
Phosphorylation
Protein Binding
Protein Kinases - metabolism
Retinaldehyde - metabolism
Rhodopsin - genetics
Rhodopsin - metabolism
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Summary:Arginine-135, located at the border between the third transmembrane domain and the second cytoplasmic loop of rhodopsin, is one of the most highly conserved amino acids in the family of G protein-coupled receptors. The effect of mutation at Arg-135 on the ability of rhodopsin to undergo desensitization was investigated. Four mutants, R135K, R135Q, R135A, and R135L, were examined for their ability to be phosphorylated by rhodopsin kinase, to bind arrestin, and to activate the rod cell G protein, transducin (Gt). All of the mutants were phosphorylated, bound arrestin, and were able to activate Gt when reconstituted with 11-cis-retinal. Surprisingly, several of the mutants could be phosphorylated by rhodopsin kinase and could bind arrestin in the absence of 11-cis-retinal but were not able to activate Gt. These observations represent the first demonstration of a mutant G protein-coupled receptor that assumes a conformation able to interact with its G protein-coupled receptor kinase and arrestin, but not with its G protein, in the absence of ligand.
ISSN:0006-2960
1520-4995
DOI:10.1021/bi9731100