title: The dynamic process of β(2)-adrenergic receptor activation.
abstract: G-protein-coupled receptors (GPCRs) can modulate diverse signaling pathways, often in a ligand-specific manner. The full range of functionally relevant GPCR conformations is poorly understood. Here, we use NMR spectroscopy to characterize the conformational dynamics of the transmembrane core of the β(2)-adrenergic receptor (β(2)AR), a prototypical GPCR. We labeled β(2)AR with (13)CH(3)ε-methionine and obtained HSQC spectra of unliganded receptor as well as receptor bound to an inverse agonist, an agonist, and a G-protein-mimetic nanobody. These studies provide evidence for conformational states not observed in crystal structures, as well as substantial conformational heterogeneity in agonist- and inverse-agonist-bound preparations. They also show that for β(2)AR, unlike rhodopsin, an agonist alone does not stabilize a fully active conformation, suggesting that the conformational link between the agonist-binding pocket and the G-protein-coupling surface is not rigid. The observed heterogeneity may be important for β(2)AR's ability to engage multiple signaling and regulatory proteins.
authors: Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA 94305, USA.
authors: Rie
authors: R
authors: Nygaard
authors: Yaozhong
authors: Y
authors: Zou
authors: Ron O
authors: RO
authors: Dror
authors: Thomas J
authors: TJ
authors: Mildorf
authors: Daniel H
authors: DH
authors: Arlow
authors: Aashish
authors: A
authors: Manglik
authors: Albert C
authors: AC
authors: Pan
authors: Corey W
authors: CW
authors: Liu
authors: Juan José
authors: JJ
authors: Fung
authors: Michael P
authors: MP
authors: Bokoch
authors: Foon Sun
authors: FS
authors: Thian
authors: Tong Sun
authors: TS
authors: Kobilka
authors: David E
authors: DE
authors: Shaw
authors: Luciano
authors: L
authors: Mueller
authors: R Scott
authors: RS
authors: Prosser
authors: Brian K
authors: BK
authors: Kobilka
mesh_terms: D058666:Adrenergic beta-2 Receptor Agonists / Q000378:metabolism; D000595:Amino Acid Sequence; D006801:Humans; D056004:Molecular Dynamics Simulation; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D011487:Protein Conformation; D018343:Receptors, Adrenergic, beta-2 / Q000737:chemistry* / Q000378:metabolism*; D015398:Signal Transduction; D013816:Thermodynamics
chemical_list: C553443:ADRB2 protein, human; D058666:Adrenergic beta-2 Receptor Agonists; D018343:Receptors, Adrenergic, beta-2
references: J Gen Physiol. 2010 Jun;135(6):555-62
references: 20513757
references: J Comput Chem. 2004 Aug;25(11):1400-15
references: 15185334
references: Biochemistry. 1996 Jun 18;35(24):7635-42
references: 8672464
references: Adv Protein Chem. 2003;63:243-90
references: 12629973
references: Nature. 2011 Jul 19;477(7366):549-55
references: 21772288
references: J Biomol NMR. 1995 Nov;6(3):277-93
references: 8520220
references: Proc Natl Acad Sci U S A. 2008 May 27;105(21):7439-44
references: 18490656
references: J Biol Chem. 1997 Jan 31;272(5):2587-90
references: 9006889
references: Proc Natl Acad Sci U S A. 2011 Nov 15;108(46):18684-9
references: 22031696
references: Nature. 2011 Mar 31;471(7340):651-5
references: 21389988
references: J Biol Chem. 1993 Mar 5;268(7):4625-36
references: 8095262
references: Proc Natl Acad Sci U S A. 2009 Jun 9;106(23):9501-6
references: 19470481
references: Nature. 2009 May 21;459(7245):356-63
references: 19458711
references: J Phys Chem B. 2010 Jun 17;114(23):7830-43
references: 20496934
references: Photochem Photobiol Sci. 2010 Nov;9(11):1466-74
references: 20967340
references: Biophys J. 2011 May 4;100(9):L47-9
references: 21539772
references: Proc Natl Acad Sci U S A. 2007 Oct 16;104(42):16657-62
references: 17925438
references: J Clin Invest. 2005 Nov;115(11):2971-4
references: 16276410
references: Physiology (Bethesda). 2010 Oct;25(5):293-303
references: 20940434
references: Proc Natl Acad Sci U S A. 2009 Mar 24;106(12):4689-94
references: 19258456
references: Nature. 2011 Jan 13;469(7329):236-40
references: 21228876
references: Trends Biochem Sci. 2009 Dec;34(12):601-11
references: 19846313
references: Prog Nucl Magn Reson Spectrosc. 2011 Feb;58(1-2):62-87
references: 21241884
references: Chembiochem. 2007 Jan 2;8(1):19-24
references: 17173267
references: Nature. 2007 Dec 13;450(7172):964-72
references: 18075575
references: Nat Chem Biol. 2011 Aug 21;7(10):692-700
references: 21857662
references: Proc Natl Acad Sci U S A. 2012 Apr 24;109(17):6733-8
references: 22493271
references: J Biomol NMR. 2010 Sep;48(1):31-47
references: 20734113
references: Biochemistry. 2004 Jul 20;43(28):8911-22
references: 15248749
references: Science. 2012 Mar 2;335(6072):1106-10
references: 22267580
references: Nature. 2007 Nov 15;450(7168):383-7
references: 17952055
references: J Biomol NMR. 2008 Jul;41(3):113-20
references: 18574698
references: J Phys Chem B. 1998 Apr 30;102(18):3586-616
references: 24889800
references: Trends Biochem Sci. 2009 Nov;34(11):540-52
references: 19836958
references: J Biol Chem. 2008 Feb 22;283(8):4967-74
references: 18063586
references: Nature. 2011 Jan 13;469(7329):175-80
references: 21228869
references: Science. 2007 Nov 23;318(5854):1266-73
references: 17962519
references: Proc Natl Acad Sci U S A. 2011 Aug 9;108(32):13118-23
references: 21778406
references: J Mol Biol. 2007 Oct 19;373(2):282-95
references: 17822711
references: Nat Commun. 2012;3:1045
references: 22948827
references: J Am Chem Soc. 2003 Aug 27;125(34):10420-8
references: 12926967
references: Nat Chem Biol. 2006 Aug;2(8):417-22
references: 16799554