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Title: Probing the remarkable thermal kinetics of visual rhodopsin with E181Q and S186A mutants

Authors:
 [1]; ORCiD logo [1];  [1];  [1];  [1];  [1];  [1];  [1];  [1]
  1. Department of Chemistry, Yale University, 225 Prospect Street, New Haven, Connecticut 06520, USA
Publication Date:
Sponsoring Org.:
USDOE
OSTI Identifier:
1365428
Grant/Contract Number:
FG02-05ER15677
Resource Type:
Journal Article: Publisher's Accepted Manuscript
Journal Name:
The Journal of Chemical Physics
Additional Journal Information:
Journal Volume: 146; Journal Issue: 21; Related Information: CHORUS Timestamp: 2017-06-22 13:10:25; Journal ID: ISSN 0021-9606
Publisher:
American Institute of Physics
Country of Publication:
United States
Language:
English

Citation Formats

Guo, Ying, Hendrickson, Heidi P., Videla, Pablo E., Chen, Ya-Na, Ho, Junming, Sekharan, Sivakumar, Batista, Victor S., Tully, John C., and Yan, Elsa C. Y. Probing the remarkable thermal kinetics of visual rhodopsin with E181Q and S186A mutants. United States: N. p., 2017. Web. doi:10.1063/1.4984818.
Guo, Ying, Hendrickson, Heidi P., Videla, Pablo E., Chen, Ya-Na, Ho, Junming, Sekharan, Sivakumar, Batista, Victor S., Tully, John C., & Yan, Elsa C. Y. Probing the remarkable thermal kinetics of visual rhodopsin with E181Q and S186A mutants. United States. doi:10.1063/1.4984818.
Guo, Ying, Hendrickson, Heidi P., Videla, Pablo E., Chen, Ya-Na, Ho, Junming, Sekharan, Sivakumar, Batista, Victor S., Tully, John C., and Yan, Elsa C. Y. Wed . "Probing the remarkable thermal kinetics of visual rhodopsin with E181Q and S186A mutants". United States. doi:10.1063/1.4984818.
@article{osti_1365428,
title = {Probing the remarkable thermal kinetics of visual rhodopsin with E181Q and S186A mutants},
author = {Guo, Ying and Hendrickson, Heidi P. and Videla, Pablo E. and Chen, Ya-Na and Ho, Junming and Sekharan, Sivakumar and Batista, Victor S. and Tully, John C. and Yan, Elsa C. Y.},
abstractNote = {},
doi = {10.1063/1.4984818},
journal = {The Journal of Chemical Physics},
number = 21,
volume = 146,
place = {United States},
year = {Wed Jun 07 00:00:00 EDT 2017},
month = {Wed Jun 07 00:00:00 EDT 2017}
}

Journal Article:
Free Publicly Available Full Text
This content will become publicly available on June 7, 2018
Publisher's Accepted Manuscript

Citation Metrics:
Cited by: 1work
Citation information provided by
Web of Science

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  • The crystal structure of rhodopsin has provided the first three-dimensional molecular model for a G-protein-coupled receptor (GPCR). Alignment of the molecular model from the crystallographic structure with the helical axes seen in cryo-electron microscopic (cryo-EM) studies provides an opportunity to investigate the properties of the molecule as a function of orientation and location within the membrane. In addition, the structure provides a starting point for modeling and rational experimental approaches of the cone pigments, the GPCRs in cone cells responsible for color vision. Homology models of the cone pigments provide a means of understanding the roles of amino acid sequencemore » differences that shift the absorption maximum of the retinal chromophore in the environments of different opsins.« less
  • Solid-state {sup 13}C NMR spectra have been obtained of bovine rhodopsin and isorhodopsin regenerated with retinal selectively {sup 13}C labeled along the polyene chain. In rhodopsin, the chemical shifts for {sup 13}C-5, {sup 13}C-6, {sup 13}C-7, {sup 13}C-14, and {sup 13}C-15 correspond closely to the chemical shifts observed in the 11-cis protonated Schiff base (PSB) model compound. Differences in chemical shift relative to the 11-cis PSB chloride salt are observed for positions 8 through 13, with the largest deshielding (6.2 ppm) localized at position 13. The localized deshielding at C-13 supports previous models of the opsin shift in rhodopsin thatmore » place a protein perturbation in the vicinity of position 13. Spectra obtained of isorhodopsin regenerated with {sup 13}C-labeled 9-cis-retinals reveal large perturbations at {sup 13}C-7 and {sup 13}C-13. The similar deshielding of the {sup 13}C-13 resonance in both pigments supports the presence of a protein perturbation near position 13. However, the chemical shifts at positions 7 and 12 in isorhodopsin are not analogous to those observed in rhodopsin and suggest that the binding site interactions near these positions are different for the two pigments. The implications of these results for the mechanism of the opsin shift in these proteins are discussed.« less
  • No abstract prepared.
  • Rhodopsin is a member of a family of receptors that contain seven transmembrane helices and are coupled to G proteins. The nature of the interactions between rhodopsin mutants and the G protein, transducin (G{sub t}), was investigated by flash photolysis in order to monitor directly G{sub t} binding and dissociation. Three mutant opsins with alterations in their cytoplasmic loops bound 11-cis-retinal to yield pigments with native rhodopsin absorption spectra, but they failed to stimulate the guanosine triphosphatase activity of G{sub t}. The opsin mutations included reversal of a charged pair conserved in all G protein-coupled receptors at the cytoplasmic bordermore » to the third transmembrane helix (mutant CD1), replacement of 13 amino acids in the second cytoplasmic loop (mutant CD2), and deletion of 13 amino acids from the third cytoplasmic loop (mutant EF1). Whereas mutant CD1 failed to bind G{sub t}, mutants CD2 and EF1 showed normal G{sub t} binding but failed to release G{sub t} in the presence of guanosine triphosphate. Therefore, it appears that at least the second and third cytoplasmic loops of rhodopsin are required for activation of bound G{sub t}.« less