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PMID: 19706523 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, U.S. Gov't, Non-P.H.S.

Structural waters define a functional channel mediating activation of the GPCR, rhodopsin.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 106 ·No. 34 ·2009-08-25 ·Pages 14367-72

Angel TE, Gupta S, Jastrzebska B, Palczewski K, Chance MR

Abstract

Structural water molecules may act as prosthetic groups indispensable for proper protein function. In the case of allosteric activation of G protein-coupled receptors (GPCRs), water likely imparts structural plasticity required for agonist-induced signal transmission. Inspection of structures of GPCR superfamily members reveals the presence of conserved embedded water molecules likely important to GPCR function. Coupling radiolytic hydroxyl radical labeling with rapid H(2)O(18) solvent mixing, we observed no exchange of these structural waters with bulk solvent in either ground state or for the Meta II or opsin states. However, the radiolysis approach permitted labeling of selected side chain residues within the transmembrane helices and revealed activation-induced changes in local structural constraints likely mediated by dynamics of both water and protein. These results suggest both a possible general mechanism for water-dependent communication in family A GPCRs based on structural conservation, and a strategy for probing membrane protein structure.

MeSH Terms
Allosteric Regulation Amino Acid Sequence Animals Binding Sites Cattle Chromatography, Liquid Hydrolysis/radiation effects Hydroxyl Radical/chemistry Mass Spectrometry/methods Models, Molecular Molecular Sequence Data Oxygen Isotopes Peptide Fragments/chemistry Protein Binding Protein Conformation Protein Structure, Secondary Protein Structure, Tertiary Receptors, G-Protein-Coupled/chemistry Rhodopsin/chemistry Solvents/chemistry Water/chemistry X-Rays
Chemicals
Oxygen Isotopes Peptide Fragments Receptors, G-Protein-Coupled Solvents Water Hydroxyl Radical Rhodopsin
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Angel Thomas E
Department of Pharmacology, Center for Proteomics and Bioinformatics, School of Medicine, Case Western Reserve University, Cleveland, OH 44106-4965, USA.
Gupta Sayan
Jastrzebska Beata
Palczewski Krzysztof
Chance Mark R
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
1091-6490
DOI
10.1073/pnas.0901074106
Published
2009-08-25
Epub
2009-00-13
Pages
14367-72
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC2732891
Subset
IM
Grants
NEI NIH HHS · R01 EY009339 · United States
NEI NIH HHS · T32 EY007157 · United States
NIGMS NIH HHS · GM079191 · United States
NIBIB NIH HHS · EB01979 · United States
NIBIB NIH HHS · P30 EB009998 · United States
NEI NIH HHS · T32EY007157 · United States
NEI NIH HHS · EY09339 · United States
NIBIB NIH HHS · P41 EB001979 · United States
NIGMS NIH HHS · R01 GM079191 · United States
NIBIB NIH HHS · EB09998 · United States
External Links
PubMed source DOI: 10.1073/pnas.0901074106 PMC Full Text View journal details
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