Home LiteratureArticle Details
PMID: 19433801 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

Conserved waters mediate structural and functional activation of family A (rhodopsin-like) G protein-coupled receptors.

Angel TE, Chance MR, Palczewski K

Abstract

G protein-coupled receptors with seven transmembrane alpha-helices (GPCRs) comprise the largest receptor superfamily and are involved in detecting a wide variety of extracellular stimuli. The availability of high-resolution crystal structures of five prototypical GPCRs, bovine and squid rhodopsin, engineered A(2A)-adenosine, beta(1)- and beta(2)-adrenergic receptors, permits comparative analysis of features common to these and likely all GPCRs. We provide an analysis of the distribution of water molecules in the transmembrane region of these GPCR structures and find conserved contacts with microdomains demonstrated to be involved in receptor activation. Colocalization of water molecules associating with highly conserved and functionally important residues in several of these GPCR crystal structures supports the notion that these waters are likely to be as important to proper receptor function as the conserved residues. Moreover, in the absence of large conformational changes in rhodopsin after photoactivation, we propose that ordered waters contribute to the functional plasticity needed to transmit activation signals from the retinal-binding pocket to the cytoplasmic face of rhodopsin and that fundamental features of the mechanism of activation, involving these conserved waters, are shared by many if not all family A receptors.

MeSH Terms
Animals Cattle Crystallography, X-Ray Decapodiformes Hydrogen Bonding Models, Molecular Protein Binding Protein Structure, Quaternary Rhodopsin/chemistry,classification,genetics,metabolism Water/chemistry,metabolism
Chemicals
Water Rhodopsin
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Angel Thomas E
Department of Pharmacology, School of Medicine, Case Western Reserve University, Cleveland, OH 44106-4965, USA.
Chance Mark R
Palczewski Krzysztof
References (59)
59 references, click to expand
  1. New G-protein-coupled receptor crystal structures: insights and limitations.
    Trends Pharmacol Sci. 2008 Feb;29(2):79-83 PMID: 18194818
  2. Clustal W and Clustal X version 2.0.
    Bioinformatics. 2007 Nov 1;23(21):2947-8 PMID: 17846036
  3. Advances in determination of a high-resolution three-dimensional structure of rhodopsin, a model of G-protein-coupled receptors (GPCRs).
    Biochemistry. 2001 Jul 3;40(26):7761-72 PMID: 11425302
  4. Evidence for a bound water molecule next to the retinal Schiff base in bacteriorhodopsin and rhodopsin: a resonance Raman study of the Schiff base hydrogen/deuterium exchange.
    Biophys J. 1994 Apr;66(4):1129-36 PMID: 8038384
  5. Constitutive activation of the beta2 adrenergic receptor alters the orientation of its sixth membrane-spanning segment.
    J Biol Chem. 1997 Jul 25;272(30):18546-9 PMID: 9228019
  6. Crystal structure of rhodopsin: A G protein-coupled receptor.
    Science. 2000 Aug 4;289(5480):739-45 PMID: 10926528
  7. Chimeric alpha 2-,beta 2-adrenergic receptors: delineation of domains involved in effector coupling and ligand binding specificity.
    Science. 1988 Jun 3;240(4857):1310-6 PMID: 2836950
  8. Hydroxyl radical-mediated modification of proteins as probes for structural proteomics.
    Chem Rev. 2007 Aug;107(8):3514-43 PMID: 17683160
  9. Protonation states of membrane-embedded carboxylic acid groups in rhodopsin and metarhodopsin II: a Fourier-transform infrared spectroscopy study of site-directed mutants.
    Proc Natl Acad Sci U S A. 1993 Nov 1;90(21):10206-10 PMID: 7901852
  10. Synergistic contributions of the functional groups of epinephrine to its affinity and efficacy at the beta2 adrenergic receptor.
    Mol Pharmacol. 2004 May;65(5):1181-90 PMID: 15102946
  11. The light reaction in the bleaching of rhodopsin.
    Science. 1950 Feb 17;111(2877):179-81 PMID: 15403120
  12. Crystal structure of opsin in its G-protein-interacting conformation.
    Nature. 2008 Sep 25;455(7212):497-502 PMID: 18818650
  13. Role of the conserved NPxxY(x)5,6F motif in the rhodopsin ground state and during activation.
    Proc Natl Acad Sci U S A. 2003 Mar 4;100(5):2290-5 PMID: 12601165
  14. Effect of protein hydration on receptor conformation: decreased levels of bound water promote metarhodopsin II formation.
    Biochemistry. 1999 Jun 15;38(24):7617-23 PMID: 10387000
  15. The CCP4 suite: programs for protein crystallography.
    Acta Crystallogr D Biol Crystallogr. 1994 Sep 1;50(Pt 5):760-3 PMID: 15299374
  16. Agonists induce conformational changes in transmembrane domains III and VI of the beta2 adrenoceptor.
    EMBO J. 1997 Nov 17;16(22):6737-47 PMID: 9362488
  17. Crystal structure of the ligand-free G-protein-coupled receptor opsin.
    Nature. 2008 Jul 10;454(7201):183-7 PMID: 18563085
  18. The role of internal water molecules in the structure and function of the rhodopsin family of G protein-coupled receptors.
    Chembiochem. 2007 Jan 2;8(1):19-24 PMID: 17173267
  19. Rhodopsin mutants discriminate sites important for the activation of rhodopsin kinase and Gt.
    J Biol Chem. 1995 Feb 3;270(5):2112-9 PMID: 7836439
  20. Role of phosphorylation in agonist-promoted beta 2-adrenergic receptor sequestration. Rescue of a sequestration-defective mutant receptor by beta ARK1.
    J Biol Chem. 1995 Oct 20;270(42):24782-9 PMID: 7559596
  21. Functional role of internal water molecules in rhodopsin revealed by X-ray crystallography.
    Proc Natl Acad Sci U S A. 2002 Apr 30;99(9):5982-7 PMID: 11972040
  22. Crystal structure of the human beta2 adrenergic G-protein-coupled receptor.
    Nature. 2007 Nov 15;450(7168):383-7 PMID: 17952055
  23. Evidence for a model of agonist-induced activation of 5-hydroxytryptamine 2A serotonin receptors that involves the disruption of a strong ionic interaction between helices 3 and 6.
    J Biol Chem. 2002 Mar 29;277(13):11441-9 PMID: 11801601
  24. Members of the G protein-coupled receptor kinase family that phosphorylate the beta2-adrenergic receptor facilitate sequestration.
    Biochemistry. 1996 Apr 2;35(13):4155-60 PMID: 8672451
  25. The hydrogen-bonding network of water molecules and the peptide backbone in the region connecting Asp83, Gly120, and Glu113 in bovine rhodopsin.
    Biochemistry. 1998 Dec 8;37(49):17216-22 PMID: 9860835
  26. Structural changes of water molecules during the photoactivation processes in bovine rhodopsin.
    Biochemistry. 2003 Aug 19;42(32):9619-25 PMID: 12911303
  27. Rhodopsin structure, dynamics, and activation: a perspective from crystallography, site-directed spin labeling, sulfhydryl reactivity, and disulfide cross-linking.
    Adv Protein Chem. 2003;63:243-90 PMID: 12629973
  28. Similar structures and shared switch mechanisms of the beta2-adrenoceptor and the parathyroid hormone receptor. Zn(II) bridges between helices III and VI block activation.
    J Biol Chem. 1999 Jun 11;274(24):17033-41 PMID: 10358054
  29. Crystal structure of squid rhodopsin.
    Nature. 2008 May 15;453(7193):363-7 PMID: 18480818
  30. How many water molecules can be detected by protein crystallography?
    Acta Crystallogr D Biol Crystallogr. 1999 Feb;55(Pt 2):479-83 PMID: 10089359
  31. Evolutionary trace of G protein-coupled receptors reveals clusters of residues that determine global and class-specific functions.
    J Biol Chem. 2004 Feb 27;279(9):8126-32 PMID: 14660595
  32. Rhodopsin activation: effects on the metarhodopsin I-metarhodopsin II equilibrium of neutralization or introduction of charged amino acids within putative transmembrane segments.
    Biochemistry. 1993 Dec 28;32(51):14176-82 PMID: 8260503
  33. Location of Trp265 in metarhodopsin II: implications for the activation mechanism of the visual receptor rhodopsin.
    J Mol Biol. 2006 Mar 17;357(1):163-72 PMID: 16414074
  34. The 2.6 angstrom crystal structure of a human A2A adenosine receptor bound to an antagonist.
    Science. 2008 Nov 21;322(5905):1211-7 PMID: 18832607
  35. Two different forms of metarhodopsin II: Schiff base deprotonation precedes proton uptake and signaling state.
    Proc Natl Acad Sci U S A. 1993 Aug 15;90(16):7849-53 PMID: 8356093
  36. Effect of ethanol and osmotic stress on receptor conformation. Reduced water activity amplifies the effect of ethanol on metarhodopsin II formation.
    J Biol Chem. 2000 Feb 25;275(8):5355-60 PMID: 10681509
  37. Crystal structure of a photoactivated deprotonated intermediate of rhodopsin.
    Proc Natl Acad Sci U S A. 2006 Oct 31;103(44):16123-8 PMID: 17060607
  38. High-resolution crystal structure of an engineered human beta2-adrenergic G protein-coupled receptor.
    Science. 2007 Nov 23;318(5854):1258-65 PMID: 17962520
  39. Agonist-induced conformational changes at the cytoplasmic side of transmembrane segment 6 in the beta 2 adrenergic receptor mapped by site-selective fluorescent labeling.
    J Biol Chem. 2001 Mar 23;276(12):9279-90 PMID: 11118431
  40. Photoactivation of rhodopsin causes an increased hydrogen-deuterium exchange of buried peptide groups.
    Biophys J. 1998 Jan;74(1):192-8 PMID: 9449322
  41. High-resolution distance mapping in rhodopsin reveals the pattern of helix movement due to activation.
    Proc Natl Acad Sci U S A. 2008 May 27;105(21):7439-44 PMID: 18490656
  42. Structure of bovine rhodopsin in a trigonal crystal form.
    J Mol Biol. 2004 Nov 5;343(5):1409-38 PMID: 15491621
  43. Water as an active constituent in cell biology.
    Chem Rev. 2008 Jan;108(1):74-108 PMID: 18095715
  44. Opsin/all-trans-retinal complex activates transducin by different mechanisms than photolyzed rhodopsin.
    Biochemistry. 1996 Mar 5;35(9):2901-8 PMID: 8608127
  45. Requirement of rigid-body motion of transmembrane helices for light activation of rhodopsin.
    Science. 1996 Nov 1;274(5288):768-70 PMID: 8864113
  46. The retinal conformation and its environment in rhodopsin in light of a new 2.2 A crystal structure.
    J Mol Biol. 2004 Sep 10;342(2):571-83 PMID: 15327956
  47. A comparison of the efficiency of G protein activation by ligand-free and light-activated forms of rhodopsin.
    Biophys J. 1997 Dec;73(6):3182-91 PMID: 9414230
  48. Characterization of the mutant visual pigment responsible for congenital night blindness: a biochemical and Fourier-transform infrared spectroscopy study.
    Biochemistry. 1996 Jun 11;35(23):7536-45 PMID: 8652533
  49. Linkage between the intramembrane H-bond network around aspartic acid 83 and the cytosolic environment of helix 8 in photoactivated rhodopsin.
    J Mol Biol. 2007 Mar 2;366(4):1129-41 PMID: 17196983
  50. Opioid receptor random mutagenesis reveals a mechanism for G protein-coupled receptor activation.
    Nat Struct Biol. 2003 Aug;10(8):629-36 PMID: 12847517
  51. Light-stable rhodopsin. II. An opsin mutant (TRP-265----Phe) and a retinal analog with a nonisomerizable 11-cis configuration form a photostable chromophore.
    J Biol Chem. 1992 Apr 5;267(10):6770-5 PMID: 1532391
  52. Structure of a beta1-adrenergic G-protein-coupled receptor.
    Nature. 2008 Jul 24;454(7203):486-91 PMID: 18594507
  53. Functional role of the "ionic lock"--an interhelical hydrogen-bond network in family A heptahelical receptors.
    J Mol Biol. 2008 Jul 18;380(4):648-55 PMID: 18554610
  54. Do we underestimate the importance of water in cell biology?
    Nat Rev Mol Cell Biol. 2006 Nov;7(11):861-6 PMID: 16955076
  55. Metarhodopsin-II stabilization by crosslinked Gtalpha C-terminal peptides and implications for the mechanism of GPCR-G protein coupling.
    Vision Res. 2006 Dec;46(27):4547-55 PMID: 17014882
  56. Structural changes in the lumirhodopsin-to-metarhodopsin I conversion of air-dried bovine rhodopsin.
    Biochemistry. 1995 Dec 26;34(51):16758-63 PMID: 8527450
  57. G protein-coupled receptor rhodopsin.
    Annu Rev Biochem. 2006;75:743-67 PMID: 16756510
  58. Effect of carboxyl mutations on functional properties of bovine rhodopsin.
    Biophys Chem. 1995 Sep-Oct;56(1-2):79-87 PMID: 7662872
  59. Ordered water molecules as key allosteric mediators in a cooperative dimeric hemoglobin.
    Proc Natl Acad Sci U S A. 1996 Dec 10;93(25):14526-31 PMID: 8962085
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
Published
2009-05-26
Epub
2009-00-11
Pages
8555-60
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC2688986
Subset
IM
Grants
NEI NIH HHS · R01 EY009339 · United States
NEI NIH HHS · T32 EY007157 · United States
NIGMS NIH HHS · GM079191 · United States
NEI NIH HHS · R01 EY009339-20 · United States
NIBIB NIH HHS · EB01979 · United States
NIGMS NIH HHS · R01 GM079191 · United States
NIBIB NIH HHS · P30 EB009998 · United States
NEI NIH HHS · T32EY007157 · United States
NEI NIH HHS · EY09339 · United States
NIGMS NIH HHS · R01 GM079191-03 · United States
NIBIB NIH HHS · P41 EB001979 · United States
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com