Rhodopsin activation blocked by metal-ion-binding sites linking transmembrane helices C and F

被引:376
作者
Sheikh, SP
Zvyaga, TA
Lichtarge, O
Sakmar, TP
Bourne, HR
机构
[1] UNIV CALIF SAN FRANCISCO,DEPT MOL & CELLULAR BIOL,SAN FRANCISCO,CA 94143
[2] UNIV CALIF SAN FRANCISCO,INST CARDIOVASC RES,SAN FRANCISCO,CA 94143
[3] ROCKEFELLER UNIV,HOWARD HUGHES MED INST,DEPT MOL BIOL & BIOCHEM,NEW YORK,NY 10021
关键词
D O I
10.1038/383347a0
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
A LARGE superfamily of receptors containing seven transmembrane (TRI) helices transmits hormonal and sensory signals across the plasma membrane to heterotrimeric G proteins at the cytoplasmic face of the membrane. To investigate how G-protein-coupled receptors work at the molecular level, we have engineered metal-ion-binding sites between TM helices to restrain activation-induced conformational change in specific locations. In rhodopsin, the photoreceptor of retinal rod cells, we substituted histidine residues for natural amino acids at the cytoplasmic ends of the TM helices C and F. The resulting mutant proteins were able to activate the visual G protein transducin in the absence but not in the presence of metal ions. These results indicate that the TM helices C and F are in close proximity and suggest that movements of these helices relative to one another are required for transducin activation. Thus a change in the orientations of TRI helices C and F is likely to be a key element in the mechanism for coupling binding of ligands (or isomerization of retinal) to the activation of G-protein-coupled receptors.
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页码:347 / 350
页数:4
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