{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T23:22:09Z","timestamp":1791328929969,"version":"4.3.1"},"reference-count":66,"publisher":"Public Library of Science (PLoS)","issue":"3","license":[{"start":{"date-parts":[[2014,3,24]],"date-time":"2014-03-24T00:00:00Z","timestamp":1395619200000},"content-version":"unspecified","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"DOI":"10.1371\/journal.pone.0092727","type":"journal-article","created":{"date-parts":[[2014,3,25]],"date-time":"2014-03-25T03:51:41Z","timestamp":1395719501000},"page":"e92727","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":174,"title":["The 2.1 \u00c5 Resolution Structure of Cyanopindolol-Bound \u03b21-Adrenoceptor Identifies an Intramembrane Na+ Ion that Stabilises the Ligand-Free Receptor"],"prefix":"10.1371","volume":"9","author":[{"given":"Jennifer L.","family":"Miller-Gallacher","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rony","family":"Nehm\u00e9","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tony","family":"Warne","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patricia C.","family":"Edwards","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gebhard F. X.","family":"Schertler","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrew G. W.","family":"Leslie","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christopher G.","family":"Tate","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2014,3,24]]},"reference":[{"key":"ref1","doi-asserted-by":"crossref","first-page":"356","DOI":"10.1038\/nature08144","article-title":"The structure and function of G-protein-coupled receptors","volume":"459","author":"DM Rosenbaum","year":"2009","journal-title":"Nature"},{"key":"ref2","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1038\/nature11896","article-title":"Molecular signatures of G-protein-coupled receptors","volume":"494","author":"AJ Venkatakrishnan","year":"2013","journal-title":"Nature"},{"key":"ref3","doi-asserted-by":"crossref","first-page":"386","DOI":"10.1016\/j.sbi.2009.07.004","article-title":"Engineering G protein-coupled receptors to facilitate their structure determination","volume":"19","author":"CG Tate","year":"2009","journal-title":"Curr Opin Struct Biol"},{"key":"ref4","doi-asserted-by":"crossref","first-page":"1258","DOI":"10.1126\/science.1150577","article-title":"High-resolution crystal structure of an engineered human beta2-adrenergic G protein-coupled receptor","volume":"318","author":"V Cherezov","year":"2007","journal-title":"Science"},{"key":"ref5","doi-asserted-by":"crossref","first-page":"8228","DOI":"10.1073\/pnas.1100185108","article-title":"Two distinct conformations of helix 6 observed in antagonist-bound structures of a {beta}1-adrenergic receptor","volume":"108","author":"R Moukhametzianov","year":"2011","journal-title":"Proc Natl Acad Sci U S A"},{"key":"ref6","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1038\/nature09648","article-title":"Structure of a nanobody-stabilized active state of the beta(2) adrenoceptor","volume":"469","author":"SG Rasmussen","year":"2011","journal-title":"Nature"},{"key":"ref7","doi-asserted-by":"crossref","first-page":"549","DOI":"10.1038\/nature10361","article-title":"Crystal structure of the beta(2) adrenergic receptor-Gs protein complex","volume":"477","author":"SG Rasmussen","year":"2011","journal-title":"Nature"},{"key":"ref8","doi-asserted-by":"crossref","first-page":"236","DOI":"10.1038\/nature09665","article-title":"Structure and function of an irreversible agonist-beta(2) adrenoceptor complex","volume":"469","author":"DM Rosenbaum","year":"2011","journal-title":"Nature"},{"key":"ref9","doi-asserted-by":"crossref","first-page":"11443","DOI":"10.1021\/ja105108q","article-title":"Conserved binding mode of human beta2 adrenergic receptor inverse agonists and antagonist revealed by X-ray crystallography","volume":"132","author":"D Wacker","year":"2010","journal-title":"J Am Chem Soc"},{"key":"ref10","doi-asserted-by":"crossref","first-page":"241","DOI":"10.1038\/nature09746","article-title":"The structural basis for agonist and partial agonist action on a beta(1)-adrenergic receptor","volume":"469","author":"T Warne","year":"2011","journal-title":"Nature"},{"key":"ref11","doi-asserted-by":"crossref","first-page":"486","DOI":"10.1038\/nature07101","article-title":"Structure of a beta1-adrenergic G-protein-coupled receptor","volume":"454","author":"T Warne","year":"2008","journal-title":"Nature"},{"key":"ref12","doi-asserted-by":"crossref","first-page":"841","DOI":"10.1016\/j.str.2012.03.014","article-title":"Crystal Structures of a Stabilized beta(1)-Adrenoceptor Bound to the Biased Agonists Bucindolol and Carvedilol","volume":"20","author":"T Warne","year":"2012","journal-title":"Structure"},{"key":"ref13","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1042\/BST20120228","article-title":"The importance of interactions with helix 5 in determining the efficacy of beta-adrenoceptor ligands","volume":"41","author":"T Warne","year":"2013","journal-title":"Biochem Soc Trans"},{"key":"ref14","doi-asserted-by":"crossref","first-page":"366","DOI":"10.1016\/S1043-9471(05)80049-7","article-title":"Integrated methods for the construction of three dimensional models and computational probing of structure function relations in G protein-coupled receptors","volume":"25","author":"JA Ballesteros","year":"1995","journal-title":"Methods Neurosci"},{"key":"ref15","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1146\/annurev.biophys.050708.133655","article-title":"Crystallizing membrane proteins for structure determination: use of lipidic mesophases","volume":"38","author":"M Caffrey","year":"2009","journal-title":"Annu Rev Biophys"},{"key":"ref16","doi-asserted-by":"crossref","first-page":"706","DOI":"10.1038\/nprot.2009.31","article-title":"Crystallizing membrane proteins using lipidic mesophases","volume":"4","author":"M Caffrey","year":"2009","journal-title":"Nat Protoc"},{"key":"ref17","doi-asserted-by":"crossref","first-page":"1266","DOI":"10.1126\/science.1150609","article-title":"GPCR engineering yields high-resolution structural insights into beta2-adrenergic receptor function","volume":"318","author":"DM Rosenbaum","year":"2007","journal-title":"Science"},{"key":"ref18","doi-asserted-by":"crossref","first-page":"1283","DOI":"10.1016\/j.str.2011.06.014","article-title":"Structure of the adenosine A(2A) receptor in complex with ZM241385 and the xanthines XAC and caffeine","volume":"19","author":"AS Dore","year":"2011","journal-title":"Structure"},{"key":"ref19","doi-asserted-by":"crossref","first-page":"6266","DOI":"10.1021\/bi300010w","article-title":"Membrane protein structure determination using crystallography and lipidic mesophases: recent advances and successes","volume":"51","author":"M Caffrey","year":"2012","journal-title":"Biochemistry"},{"key":"ref20","doi-asserted-by":"crossref","first-page":"298","DOI":"10.1016\/j.jmb.2011.03.075","article-title":"Thermostabilisation of an agonist-bound conformation of the human adenosine A(2A) receptor","volume":"409","author":"G Lebon","year":"2011","journal-title":"J Mol Biol"},{"key":"ref21","doi-asserted-by":"crossref","first-page":"10744","DOI":"10.1073\/pnas.0804396105","article-title":"Co-evolving stability and conformational homogeneity of the human adenosine A2a receptor","volume":"105","author":"F Magnani","year":"2008","journal-title":"Proc Natl Acad Sci U S A"},{"key":"ref22","doi-asserted-by":"crossref","first-page":"877","DOI":"10.1073\/pnas.0711253105","article-title":"Conformational thermostabilization of the beta1-adrenergic receptor in a detergent-resistant form","volume":"105","author":"MJ Serrano-Vega","year":"2008","journal-title":"Proc Natl Acad Sci U S A"},{"key":"ref23","doi-asserted-by":"crossref","first-page":"1293","DOI":"10.1016\/j.bbamem.2013.01.008","article-title":"Optimising the combination of thermostabilising mutations in the neurotensin receptor for structure determination","volume":"1828","author":"Y Shibata","year":"2013","journal-title":"Biochim Biophys Acta"},{"key":"ref24","doi-asserted-by":"crossref","first-page":"262","DOI":"10.1016\/j.jmb.2009.04.068","article-title":"Thermostabilization of the neurotensin receptor NTS1","volume":"390","author":"Y Shibata","year":"2009","journal-title":"J Mol Biol"},{"key":"ref25","doi-asserted-by":"crossref","first-page":"628","DOI":"10.1016\/j.jmb.2011.08.057","article-title":"Engineering an ultra-thermostable beta(1)-adrenoceptor","volume":"413","author":"JL Miller","year":"2011","journal-title":"J Mol Biol"},{"key":"ref26","doi-asserted-by":"crossref","first-page":"204","DOI":"10.1016\/j.pep.2009.01.014","article-title":"Development and crystallization of a minimal thermostabilized G protein-coupled receptor","volume":"65","author":"T Warne","year":"2009","journal-title":"Protein Expr Purif"},{"key":"ref27","volume":"8","unstructured":"Gaisford W, Schertler GF, Edwards P (2011) mosquito LCP: Making membrane protein crystallization accesible to the research scientist. Nature Methods <volume>8<\/volume>: : i\u2013ii."},{"key":"ref28","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1107\/S0907444905039107","article-title":"The integration of macromolecular diffraction data","volume":"62","author":"AG Leslie","year":"2006","journal-title":"Acta Crystallogr D Biol Crystallogr"},{"key":"ref29","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1107\/S0907444905036693","article-title":"Scaling and assessment of data quality","volume":"62","author":"P Evans","year":"2006","journal-title":"Acta Crystallogr D Biol Crystallogr"},{"key":"ref30","doi-asserted-by":"crossref","first-page":"658","DOI":"10.1107\/S0021889807021206","article-title":"Phaser crystallographic software","volume":"40","author":"AJ McCoy","year":"2007","journal-title":"J Appl Crystallogr"},{"key":"ref31","doi-asserted-by":"crossref","first-page":"240","DOI":"10.1107\/S0907444996012255","article-title":"Refinement of macromolecular structures by the maximum-likelihood method","volume":"53","author":"GN Murshudov","year":"1997","journal-title":"Acta Crystallogr D Biol Crystallogr"},{"key":"ref32","doi-asserted-by":"crossref","first-page":"2126","DOI":"10.1107\/S0907444904019158","article-title":"Coot: model-building tools for molecular graphics","volume":"60","author":"P Emsley","year":"2004","journal-title":"Acta Crystallogr D Biol Crystallogr"},{"key":"ref33","doi-asserted-by":"crossref","first-page":"W375","DOI":"10.1093\/nar\/gkm216","article-title":"MolProbity: all-atom contacts and structure validation for proteins and nucleic acids","volume":"35","author":"IW Davis","year":"2007","journal-title":"Nucleic Acids Res"},{"key":"ref34","doi-asserted-by":"crossref","first-page":"248","DOI":"10.1016\/0003-2697(76)90527-3","article-title":"A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding","volume":"72","author":"MM Bradford","year":"1976","journal-title":"Anal Biochem"},{"key":"ref35","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1016\/S0005-2736(02)00716-2","article-title":"Expression and purification of truncated, non-glycosylated turkey beta-adrenergic receptors for crystallization","volume":"1610","author":"T Warne","year":"2003","journal-title":"Biochim Biophys Acta"},{"key":"ref36","doi-asserted-by":"crossref","first-page":"482","DOI":"10.1016\/j.sbi.2012.03.007","article-title":"Agonist-bound structures of G protein-coupled receptors","volume":"22","author":"G Lebon","year":"2012","journal-title":"Current Opin Struct Biol"},{"key":"ref37","doi-asserted-by":"crossref","first-page":"897","DOI":"10.1016\/j.str.2008.05.001","article-title":"A specific cholesterol binding site is established by the 2.8 A structure of the human beta2-adrenergic receptor","volume":"16","author":"MA Hanson","year":"2008","journal-title":"Structure"},{"key":"ref38","doi-asserted-by":"crossref","first-page":"232","DOI":"10.1126\/science.1219218","article-title":"Structural basis for allosteric regulation of GPCRs by sodium ions","volume":"337","author":"W Liu","year":"2012","journal-title":"Science"},{"key":"ref39","doi-asserted-by":"crossref","first-page":"872","DOI":"10.1107\/S0907444902003712","article-title":"Metal-ligand geometry relevant to proteins and in proteins: sodium and potassium","volume":"58","author":"MM Harding","year":"2002","journal-title":"Acta Crystallogr D Biol Crystallogr"},{"key":"ref40","doi-asserted-by":"crossref","first-page":"669","DOI":"10.1016\/S0006-2952(00)00360-9","article-title":"Allosteric modulation of A(2A) adenosine receptors by amiloride analogues and sodium ions","volume":"60","author":"ZG Gao","year":"2000","journal-title":"Biochem Pharmacol"},{"key":"ref41","doi-asserted-by":"crossref","first-page":"1211","DOI":"10.1126\/science.1164772","article-title":"The 2.6 angstrom crystal structure of a human A2A adenosine receptor bound to an antagonist","volume":"322","author":"VP Jaakola","year":"2008","journal-title":"Science"},{"key":"ref42","doi-asserted-by":"crossref","first-page":"124","DOI":"10.1007\/BF00506514","article-title":"Sodium regulation of agonist and antagonist binding to beta-adrenoceptors in intact and Ns-deficient membranes","volume":"333","author":"M Minuth","year":"1986","journal-title":"Naunyn Schmiedebergs Arch Pharmacol"},{"key":"ref43","doi-asserted-by":"crossref","first-page":"4052","DOI":"10.1016\/S0021-9258(18)68888-X","article-title":"Site-directed mutagenesis and continuous expression of human beta-adrenergic receptors. Identification of a conserved aspartate residue involved in agonist binding and receptor activation","volume":"263","author":"FZ Chung","year":"1988","journal-title":"J Biol Chem"},{"key":"ref44","doi-asserted-by":"crossref","first-page":"10267","DOI":"10.1016\/S0021-9258(19)81509-0","article-title":"Conserved aspartic acid residues 79 and 113 of the beta-adrenergic receptor have different roles in receptor function","volume":"263","author":"CD Strader","year":"1988","journal-title":"J Biol Chem"},{"key":"ref45","doi-asserted-by":"crossref","first-page":"167","DOI":"10.1254\/jphs.FP0060640","article-title":"Beta-blockers show inverse agonism to a novel constitutively active mutant of beta1-adrenoceptor","volume":"102","author":"M Ahmed","year":"2006","journal-title":"J Pharmacol Sci"},{"key":"ref46","doi-asserted-by":"crossref","first-page":"1688","DOI":"10.1111\/j.1476-5381.2011.01629.x","article-title":"Lifting the lid on GPCRs: the role of extracellular loops","volume":"165","author":"M Wheatley","year":"2012","journal-title":"Br J Pharmacol"},{"key":"ref47","doi-asserted-by":"crossref","first-page":"1898","DOI":"10.1021\/jm201376w","article-title":"Discovery of 1,2,4-triazine derivatives as adenosine A(2A) antagonists using structure based drug design","volume":"55","author":"M Congreve","year":"2012","journal-title":"J Med Chem"},{"key":"ref48","doi-asserted-by":"crossref","first-page":"249","DOI":"10.1016\/j.tips.2012.02.005","article-title":"New insights from structural biology into the druggability of G protein-coupled receptors","volume":"33","author":"JS Mason","year":"2012","journal-title":"Trends Pharmacol Sci"},{"key":"ref49","doi-asserted-by":"crossref","first-page":"387","DOI":"10.1038\/nature11701","article-title":"High-resolution crystal structure of human protease-activated receptor 1","volume":"492","author":"C Zhang","year":"2012","journal-title":"Nature"},{"key":"ref50","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1089\/dna.1992.11.1","article-title":"Sequence alignment of the G-protein coupled receptor superfamily","volume":"11","author":"WC Probst","year":"1992","journal-title":"DNA Cell Biol"},{"key":"ref51","doi-asserted-by":"crossref","first-page":"1635","DOI":"10.1016\/S0026-895X(25)09624-5","article-title":"Site-directed mutagenesis of the human A1 adenosine receptor: influences of acidic and hydroxy residues in the first four transmembrane domains on ligand binding","volume":"50","author":"H Barbhaiya","year":"1996","journal-title":"Mol Pharmacol"},{"key":"ref52","doi-asserted-by":"crossref","first-page":"840","DOI":"10.1016\/S0026-895X(25)09700-7","article-title":"Site-directed mutagenesis of m1 muscarinic acetylcholine receptors: conserved aspartic acids play important roles in receptor function","volume":"36","author":"CM Fraser","year":"1989","journal-title":"Mol Pharmacol"},{"key":"ref53","doi-asserted-by":"crossref","first-page":"23055","DOI":"10.1016\/S0021-9258(19)49424-6","article-title":"A single residue, aspartic acid 95, in the delta opioid receptor specifies selective high affinity agonist binding","volume":"268","author":"H Kong","year":"1993","journal-title":"J Biol Chem"},{"key":"ref54","doi-asserted-by":"crossref","first-page":"968","DOI":"10.1006\/bbrc.1996.0341","article-title":"Mutation of an aspartate at position 63 in the human platelet-activating factor receptor augments binding affinity but abolishes G-protein-coupling and inositol phosphate production","volume":"219","author":"JL Parent","year":"1996","journal-title":"Biochem Biophys Res Commun"},{"key":"ref55","doi-asserted-by":"crossref","first-page":"929","DOI":"10.1124\/mol.59.4.929","article-title":"The role of a conserved inter-transmembrane domain interface in regulating alpha(2a)-adrenergic receptor conformational stability and cell-surface turnover","volume":"59","author":"MH Wilson","year":"2001","journal-title":"Mol Pharmacol"},{"key":"ref56","doi-asserted-by":"crossref","first-page":"5133","DOI":"10.1073\/pnas.90.11.5133","article-title":"Mutation of Asp74 of the rat angiotensin II receptor confers changes in antagonist affinities and abolishes G-protein coupling","volume":"90","author":"C Bihoreau","year":"1993","journal-title":"Proc Natl Acad Sci U S A"},{"key":"ref57","doi-asserted-by":"crossref","first-page":"380","DOI":"10.1016\/S0026-895X(25)13182-9","article-title":"Mutation of an aspartate at residue 89 in somatostatin receptor subtype 2 prevents Na+ regulation of agonist binding but does not alter receptor-G protein association","volume":"44","author":"H Kong","year":"1993","journal-title":"Mol Pharmacol"},{"key":"ref58","doi-asserted-by":"crossref","first-page":"552","DOI":"10.1124\/mol.108.045054","article-title":"Mutational analysis of the conserved Asp2.50 and ERY motif reveals signaling bias of the urotensin II receptor","volume":"74","author":"CD Proulx","year":"2008","journal-title":"Mol Pharmacol"},{"key":"ref59","doi-asserted-by":"crossref","first-page":"977","DOI":"10.1126\/science.1354394","article-title":"A point mutation of the alpha 2-adrenoceptor that blocks coupling to potassium but not calcium currents","volume":"257","author":"A Surprenant","year":"1992","journal-title":"Science"},{"key":"ref60","doi-asserted-by":"crossref","first-page":"651","DOI":"10.1016\/S0022-3565(24)37469-5","article-title":"Mutation of a highly conserved aspartate residue in the second transmembrane domain of the cannabinoid receptors, CB1 and CB2, disrupts G-protein coupling","volume":"285","author":"Q Tao","year":"1998","journal-title":"J Pharmacol Exp Ther"},{"key":"ref61","doi-asserted-by":"crossref","first-page":"733","DOI":"10.1016\/S0026-895X(25)11081-X","article-title":"Pivotal role for aspartate-80 in the regulation of dopamine D2 receptor affinity for drugs and inhibition of adenylyl cyclase","volume":"39","author":"KA Neve","year":"1991","journal-title":"Mol Pharmacol"},{"key":"ref62","doi-asserted-by":"crossref","first-page":"882","DOI":"10.4158\/EP10154.RA","article-title":"Hyponatremia: mechanisms and newer treatments","volume":"16","author":"ML Zeidel","year":"2010","journal-title":"Endocr Pract"},{"key":"ref63","doi-asserted-by":"crossref","first-page":"508","DOI":"10.1038\/nature11558","article-title":"Structure of the agonist-bound neurotensin receptor","volume":"490","author":"JF White","year":"2012","journal-title":"Nature"},{"key":"ref64","doi-asserted-by":"crossref","first-page":"521","DOI":"10.1038\/nature10136","article-title":"Agonist-bound adenosine A(2A) receptor structures reveal common features of GPCR activation","volume":"474","author":"G Lebon","year":"2011","journal-title":"Nature"},{"key":"ref65","doi-asserted-by":"crossref","first-page":"322","DOI":"10.1126\/science.1202793","article-title":"Structure of an agonist-bound human A2A adenosine receptor","volume":"332","author":"F Xu","year":"2011","journal-title":"Science"},{"key":"ref66","doi-asserted-by":"crossref","first-page":"3099","DOI":"10.1016\/0006-2952(73)90196-2","article-title":"Relationship between the inhibition constant (K1) and the concentration of inhibitor which causes 50 per cent inhibition (I50) of an enzymatic reaction","volume":"22","author":"Y Cheng","year":"1973","journal-title":"Biochem Pharmacol"}],"container-title":["PLoS ONE"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/dx.plos.org\/10.1371\/journal.pone.0092727","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,5,2]],"date-time":"2025-05-02T04:24:15Z","timestamp":1746159855000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0092727"}},"subtitle":[],"editor":[{"given":"Hendrik W.","family":"van Veen","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2014,3,24]]},"references-count":66,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2014,3,24]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0092727","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2014,3,24]]}}}