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PMID: 20512139 Published · ppublish English Journal Article Research Support, N.I.H., Intramural

Structural basis of G protein-coupled receptor-G protein interactions.

Nature chemical biology ·Vol. 6 ·No. 7 ·2010-07-00 ·Pages 541-8

Hu J, Wang Y, Zhang X, Lloyd JR, Li JH, Karpiak J, Costanzi S, Wess J

Abstract

The interaction of G protein-coupled receptors (GPCRs) with heterotrimeric G proteins represents one of the most fundamental biological processes. However, the molecular architecture of the GPCR-G protein complex remains poorly defined. In the present study, we applied a comprehensive GPCR-G protein alpha subunit (Galpha) chemical cross-linking strategy to map a receptor-Galpha interface, both before and after agonist-induced receptor activation. Using the M(3) muscarinic acetylcholine receptor (M3R)-Galpha(q) system as a model system, we examined the ability of approximately 250 combinations of cysteine-substituted M3R and Galpha(q) proteins to undergo cross-link formation. We identified many specific M3R-Galpha(q) contact sites, in both the inactive and active receptor conformations, allowing us to draw conclusions regarding the basic architecture of the M3R-Galpha(q) interface and the nature of the conformational changes following receptor activation. As heterotrimeric G proteins as well as most GPCRs share a high degree of structural homology, our findings should be of broad general relevance.

MeSH Terms
Amino Acid Sequence Amino Acid Substitution GTP-Binding Protein alpha Subunits/chemistry,genetics,metabolism Immunoprecipitation Mass Spectrometry Models, Molecular Mutation Protein Binding Protein Conformation Receptors, Muscarinic/chemistry,genetics,metabolism
Chemicals
GTP-Binding Protein alpha Subunits Receptors, Muscarinic
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Hu Jianxin
Molecular Signaling Section, Laboratory of Bioorganic Chemistry, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland, USA. jianxinh@niddk.nih.gov
Wang Yan
Zhang Xiaohong
Lloyd John R
Li Jian Hua
Karpiak Joel
Costanzi Stefano
Wess Jürgen
References (46)
46 references, click to expand
  1. Receptor-mediated activation of heterotrimeric G-proteins: current structural insights.
    Mol Pharmacol. 2007 Aug;72(2):219-30 PMID: 17430994
  2. The ants go marching two by two: oligomeric structure of G-protein-coupled receptors.
    Mol Pharmacol. 2004 Nov;66(5):1077-82 PMID: 15319448
  3. Role of the PAR1 receptor 8th helix in signaling: the 7-8-1 receptor activation mechanism.
    J Biol Chem. 2006 Feb 17;281(7):4109-16 PMID: 16354660
  4. Molecular basis of receptor/G-protein-coupling selectivity.
    Pharmacol Ther. 1998 Dec;80(3):231-64 PMID: 9888696
  5. Heterotrimeric G protein activation by G-protein-coupled receptors.
    Nat Rev Mol Cell Biol. 2008 Jan;9(1):60-71 PMID: 18043707
  6. Protein complexes involved in heptahelical receptor-mediated signal transduction.
    Recept Channels. 2003;9(3):169-94 PMID: 12775338
  7. Structural evidence for a sequential release mechanism for activation of heterotrimeric G proteins.
    J Mol Biol. 2009 Nov 6;393(4):882-97 PMID: 19703466
  8. Crystal structure of opsin in its G-protein-interacting conformation.
    Nature. 2008 Sep 25;455(7212):497-502 PMID: 18818650
  9. Mapping of contact sites in complex formation between transducin and light-activated rhodopsin by covalent crosslinking: use of a photoactivatable reagent.
    Proc Natl Acad Sci U S A. 2001 Apr 24;98(9):4877-82 PMID: 11320237
  10. Molecular biology of muscarinic acetylcholine receptors.
    Crit Rev Neurobiol. 1996;10(1):69-99 PMID: 8853955
  11. Mechanism of the receptor-catalyzed activation of heterotrimeric G proteins.
    Nat Struct Mol Biol. 2006 Sep;13(9):772-7 PMID: 16892066
  12. Site of G protein binding to rhodopsin mapped with synthetic peptides from the alpha subunit.
    Science. 1988 Aug 12;241(4867):832-5 PMID: 3136547
  13. Seven-transmembrane receptors.
    Nat Rev Mol Cell Biol. 2002 Sep;3(9):639-50 PMID: 12209124
  14. Use of a disulfide cross-linking strategy to study muscarinic receptor structure and mechanisms of activation.
    J Biol Chem. 1999 Jun 4;274(23):16629-40 PMID: 10347230
  15. Crystal structure of the ligand-free G-protein-coupled receptor opsin.
    Nature. 2008 Jul 10;454(7201):183-7 PMID: 18563085
  16. Optical techniques to analyze real-time activation and signaling of G-protein-coupled receptors.
    Trends Pharmacol Sci. 2008 Mar;29(3):159-65 PMID: 18262662
  17. Disease-causing mutation in GPR54 reveals the importance of the second intracellular loop for class A G-protein-coupled receptor function.
    J Biol Chem. 2008 Nov 7;283(45):31068-78 PMID: 18772143
  18. Distinct structural changes in a G protein-coupled receptor caused by different classes of agonist ligands.
    J Biol Chem. 2007 Sep 7;282(36):26284-93 PMID: 17623649
  19. Crystal structure of the human beta2 adrenergic G-protein-coupled receptor.
    Nature. 2007 Nov 15;450(7168):383-7 PMID: 17952055
  20. Real-time monitoring of receptor and G-protein interactions in living cells.
    Nat Methods. 2005 Mar;2(3):177-84 PMID: 15782186
  21. Anatomical profiling of G protein-coupled receptor expression.
    Cell. 2008 Oct 31;135(3):561-71 PMID: 18984166
  22. 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
  23. Pharmacology of a constitutively active muscarinic receptor generated by random mutagenesis.
    J Pharmacol Exp Ther. 1995 Dec;275(3):1274-9 PMID: 8531092
  24. 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
  25. Palmitoylation but not the extreme amino-terminus of Gq alpha is required for coupling to the NK2 receptor.
    FEBS Lett. 1994 Nov 7;354(2):195-9 PMID: 7957923
  26. Substitution of three amino acids switches receptor specificity of Gq alpha to that of Gi alpha.
    Nature. 1993 May 20;363(6426):274-6 PMID: 8387644
  27. Hydrophobic amino acid in the i2 loop plays a key role in receptor-G protein coupling.
    J Biol Chem. 1993 Oct 25;268(30):22273-6 PMID: 8226735
  28. Crystal structure of rhodopsin: A G protein-coupled receptor.
    Science. 2000 Aug 4;289(5480):739-45 PMID: 10926528
  29. The transitory complex between photoexcited rhodopsin and transducin. Reciprocal interaction between the retinal site in rhodopsin and the nucleotide site in transducin.
    Eur J Biochem. 1989 Oct 1;184(3):687-98 PMID: 2509200
  30. High-resolution crystal structure of an engineered human beta2-adrenergic G protein-coupled receptor.
    Science. 2007 Nov 23;318(5854):1258-65 PMID: 17962520
  31. Insights into G protein structure, function, and regulation.
    Endocr Rev. 2003 Dec;24(6):765-81 PMID: 14671004
  32. Characterization of the residues in helix 8 of the human beta1-adrenergic receptor that are involved in coupling the receptor to G proteins.
    J Biol Chem. 2006 May 5;281(18):12896-907 PMID: 16500896
  33. 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
  34. How do receptors activate G proteins?
    Adv Protein Chem. 2007;74:67-93 PMID: 17854655
  35. Coupling of the alpha 2A-adrenergic receptor to multiple G-proteins. A simple approach for estimating receptor-G-protein coupling efficiency in a transient expression system.
    J Biol Chem. 1994 Feb 25;269(8):5730-4 PMID: 7907086
  36. Palmitoylation is required for signaling functions and membrane attachment of Gq alpha and Gs alpha.
    J Biol Chem. 1993 Nov 25;268(33):25001-8 PMID: 8227063
  37. Methods to monitor the quaternary structure of G protein-coupled receptors.
    FEBS J. 2005 Jun;272(12):2914-25 PMID: 15955052
  38. Mutation of the fourth cytoplasmic loop of rhodopsin affects binding of transducin and peptides derived from the carboxyl-terminal sequences of transducin alpha and gamma subunits.
    J Biol Chem. 2000 Jan 21;275(3):1937-43 PMID: 10636895
  39. Heterotrimeric G proteins precouple with G protein-coupled receptors in living cells.
    Proc Natl Acad Sci U S A. 2005 Dec 20;102(51):18706-11 PMID: 16352729
  40. Structure of a beta1-adrenergic G-protein-coupled receptor.
    Nature. 2008 Jul 24;454(7203):486-91 PMID: 18594507
  41. Mapping of contact sites in complex formation between light-activated rhodopsin and transducin by covalent crosslinking: use of a chemically preactivated reagent.
    Proc Natl Acad Sci U S A. 2001 Apr 24;98(9):4883-7 PMID: 11320238
  42. Single-cysteine substitution mutants at amino acid positions 306-321 in rhodopsin, the sequence between the cytoplasmic end of helix VII and the palmitoylation sites: sulfhydryl reactivity and transducin activation reveal a tertiary structure.
    Biochemistry. 1999 Jun 22;38(25):7925-30 PMID: 10387034
  43. Active Galpha(q) subunits and M3 acetylcholine receptors promote distinct modes of association of RGS2 with the plasma membrane.
    FEBS Lett. 2007 Feb 20;581(4):764-70 PMID: 17275815
  44. Structural basis for nucleotide exchange on G alpha i subunits and receptor coupling specificity.
    Proc Natl Acad Sci U S A. 2007 Feb 6;104(6):2001-6 PMID: 17264214
  45. Structural diversity of G protein-coupled receptors and significance for drug discovery.
    Nat Rev Drug Discov. 2008 Apr;7(4):339-57 PMID: 18382464
  46. Receptor and betagamma binding sites in the alpha subunit of the retinal G protein transducin.
    Science. 1997 Jan 17;275(5298):381-4 PMID: 8994033
Article Info
Journal
Nature chemical biology
Abbr.
Nat Chem Biol
ISSN
1552-4469
DOI
10.1038/nchembio.385
Email
jianxinh@niddk.nih.gov
Published
2010-07-00
Epub
2010-00-30
Pages
541-8
Language
English
Region
United States
NLM ID
101231976
PMCID
PMC3104732
Subset
IM
Grants
Intramural NIH HHS · Z01 DK031129-01 · United States
Intramural NIH HHS · Z01 DK031129-02 · United States
External Links
PubMed source DOI: 10.1038/nchembio.385 PMC Full Text View journal details
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