Publication Date:
2010-04-13
Description:
Rhodopsin is a prototypical heptahelical family A G-protein-coupled receptor (GPCR) responsible for dim-light vision. Light isomerizes rhodopsin's retinal chromophore and triggers concerted movements of transmembrane helices, including an outward tilting of helix 6 (H6) and a smaller movement of H5, to create a site for G-protein binding and activation. However, the precise temporal sequence and mechanism underlying these helix rearrangements is unclear. We used site-directed non-natural amino acid mutagenesis to engineer rhodopsin with p-azido-l-phenylalanine residues incorporated at selected sites, and monitored the azido vibrational signatures using infrared spectroscopy as rhodopsin proceeded along its activation pathway. Here we report significant changes in electrostatic environments of the azido probes even in the inactive photoproduct Meta I, well before the active receptor state was formed. These early changes suggest a significant rotation of H6 and movement of the cytoplasmic part of H5 away from H3. Subsequently, a large outward tilt of H6 leads to opening of the cytoplasmic surface to form the active receptor photoproduct Meta II. Thus, our results reveal early conformational changes that precede larger rigid-body helix movements, and provide a basis to interpret recent GPCR crystal structures and to understand conformational sub-states observed during the activation of other GPCRs.〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Ye, Shixin -- Zaitseva, Ekaterina -- Caltabiano, Gianluigi -- Schertler, Gebhard F X -- Sakmar, Thomas P -- Deupi, Xavier -- Vogel, Reiner -- England -- Nature. 2010 Apr 29;464(7293):1386-9. doi: 10.1038/nature08948. Epub 2010 Apr 11.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Laboratory of Molecular Biology and Biochemistry, The Rockefeller University, 1230 York Avenue, New York, New York 10065, USA.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/20383122" target="_blank"〉PubMed〈/a〉
Keywords:
Azides/analysis/*metabolism/radiation effects
;
Cell Line
;
Humans
;
*Infrared Rays
;
Models, Molecular
;
Movement
;
Phenylalanine/*analogs & derivatives/analysis/genetics/metabolism/radiation
;
effects
;
Protein Conformation
;
Rhodopsin/chemistry/*genetics/*metabolism
;
Spectroscopy, Fourier Transform Infrared
;
Static Electricity
;
Vibration
Print ISSN:
0028-0836
Electronic ISSN:
1476-4687
Topics:
Biology
,
Chemistry and Pharmacology
,
Medicine
,
Natural Sciences in General
,
Physics
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