期刊
JOURNAL OF MOLECULAR BIOLOGY
卷 372, 期 1, 页码 50-66出版社
ACADEMIC PRESS LTD ELSEVIER SCIENCE LTD
DOI: 10.1016/j.jmb.2007.03.046
关键词
G protein-coupled receptors; membranes; solid-state NMR; retinal; rhodopsin
资金
- NEI NIH HHS [R01 EY012049, EY 12049] Funding Source: Medline
- NIGMS NIH HHS [R01 GM036564, GM 36564] Funding Source: Medline
Rhodopsin is a prototype for G protein-coupled receptors (GPCRs) that are implicated in many biological responses in humans. A site-directed H-2 NMR approach was used for structural analysis of retinal within its binding cavity in the dark and pre-activated meta I states. Retinal was H-2 NMR and was used to regenerate the opsin apoprotein. Solid-state spectra were acquired for aligned membranes in the low-temperature lipid gel phase versus the tilt angle to the magnetic field. Data reduction assumed a static uniaxial distribution, and gave the retinylidene methyl bond orientations plus the alignment disorder (mosaic spread). The dark-state H-2 NMR structure of 11-cis-retinal shows torsional twisting of the polyene chain and the beta-ionone ring. The ligand undergoes restricted motion, as evinced by order parameters of approximate to 0.9 for the spinning C-(CH3)-H-2 groups, with off-axial fluctuations of approximate to 15 degrees. Retinal is accommodated within the rhodopsin binding pocket with a negative pre-twist about the C11=C12 double bond that explains its rapid photochemistry and the trajectory of 11-cis to trans isomerization. In the cryo-trapped meta I state, the H-2 NMR structure shows a reduction of the polyene strain, while torsional twisting of the [beta-ionone ring is maintained. Distortion of the retinal conformation is interpreted through substituent control of receptor activation. Steric hindrance between trans retinal and Trp265 can trigger formation of the subsequent activated meta II state. Our results are pertinent to quantum and molecular mechanics simulations of ligands bound to GPCRs, and illustrate how H-2 NMR can be applied to study their biological mechanisms of action.(c) 2007 Elsevier Ltd. All rights reserved.
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