Activation mechanism of endothelin ETB receptor by endothelin-1

Endothelin, a 21-amino-acid peptide, participates in various physiological processes, such as regulation of vascular tone, humoral homeostasis, neural crest cell development and neurotransmission. Endothelin and its G-protein-coupled receptor are involved in the development of various diseases, such as pulmonary arterial hypertension, and thus are important therapeutic targets. Here we report crystal structures of human endothelin type B receptor in the ligand-free form and in complex with the endogenous agonist endothelin-1. The structures and mutation analysis reveal the mechanism for the isopeptide selectivity between endothelin-1 and -3. Transmembrane helices 1, 2, 6 and 7 move and envelop the entire endothelin peptide, in a virtually irreversible manner. The agonist-induced conformational changes are propagated to the receptor core and the cytoplasmic G-protein coupling interface, and probably induce conformational flexibility in TM6. A comparison with the M2 muscarinic receptor suggests a shared mechanism for signal transduction in class A G-protein-coupled receptors.

[1]  T. Watanabe,et al.  Human endothelin receptor ETB. Amino acid sequence requirements for super stable complex formation with its ligand. , 1994, The Journal of biological chemistry.

[2]  Xavier Robert,et al.  Deciphering key features in protein structures with the new ENDscript server , 2014, Nucleic Acids Res..

[3]  P. Vigne,et al.  Aspirin and sodium salicylate inhibit endothelin ETA receptors by an allosteric type of mechanism. , 2000, Molecular pharmacology.

[4]  M. Babu,et al.  Molecular signatures of G-protein-coupled receptors , 2013, Nature.

[5]  Randy J. Read,et al.  Phaser crystallographic software , 2007, Journal of applied crystallography.

[6]  C. E. Peishoff,et al.  Lysine 182 of endothelin B receptor modulates agonist selectivity and antagonist affinity: evidence for the overlap of peptide and non-peptide ligand binding sites. , 1994, Biochemistry.

[7]  B. Krumm,et al.  Structural prerequisites for G-protein activation by the neurotensin receptor , 2015, Nature Communications.

[8]  A. Davenport,et al.  Endothelin@25 – new agonists, antagonists, inhibitors and emerging research frontiers: IUPHAR Review 12 , 2014, British journal of pharmacology.

[9]  Y. Hiroaki,et al.  Interactions of endothelin receptor subtypes A and B with Gi, Go, and Gq in reconstituted phospholipid vesicles. , 1999, Biochemistry.

[10]  R. Kedzierski,et al.  Endothelin system: the double-edged sword in health and disease. , 2001, Annual review of pharmacology and toxicology.

[11]  H. Tabuchi,et al.  Purification of an endothelin receptor from human placenta. , 1990, Biochemical and biophysical research communications.

[12]  A. Ergul,et al.  Identification of receptor binding and activation sites in endothelin-1 by use of site-directed mutagenesis. , 1995, Circulation research.

[13]  J. Wess,et al.  Activation and allosteric modulation of a muscarinic acetylcholine receptor , 2013, Nature.

[14]  A. Dilella,et al.  Molecular cloning and characterization of the major endothelin receptor subtype in porcine cerebellum. , 1992, Molecular pharmacology.

[15]  G. Krause,et al.  Structural determinants for selective recognition of peptide ligands for endothelin receptor subtypes ETA and ETB , 2009, Journal of peptide science : an official publication of the European Peptide Society.

[16]  J. Shiloach,et al.  Structure of the agonist-bound neurotensin receptor , 2012, Nature.

[17]  P. Emsley,et al.  Features and development of Coot , 2010, Acta crystallographica. Section D, Biological crystallography.

[18]  M. Yanagisawa,et al.  Endothelin: 20 years from discovery to therapy. , 2008, Canadian journal of physiology and pharmacology.

[19]  M. Fujino,et al.  Endothelin-1 analogues substituted at both position 18 and 19: highly potent endothelin antagonists with no selectivity for either receptor subtype ETA or ETB. , 1993, Journal of medicinal chemistry.

[20]  J. Tam,et al.  D-amino acid scan of endothelin: importance of amino acids adjacent to cysteinyl residues in isomeric selectivity. , 1995, Peptide research.

[21]  Robert W. Janes,et al.  The crystal structure of human endothelin , 1994, Nature Structural Biology.

[22]  V. Seifert,et al.  Ambrisentan, a non-peptide endothelin receptor antagonist. , 2006, Cardiovascular drug reviews.

[23]  W. Neidhart,et al.  Pathophysiological role of endothelin revealed by the first orally active endothelin receptor antagonist , 1993, Nature.

[24]  Anthony P. Davenport,et al.  International Union of Pharmacology. XXIX. Update on Endothelin Receptor Nomenclature , 2002, Pharmacological Reviews.

[25]  Yoko Shibata,et al.  Conformational thermostabilization of the β1-adrenergic receptor in a detergent-resistant form , 2008, Proceedings of the National Academy of Sciences.

[26]  A. Sakamoto,et al.  Palmitoylation of Human EndothelinB , 1997, The Journal of Biological Chemistry.

[27]  J. Tam,et al.  Alanine scan of endothelin: Importance of aromatic residues , 1994, Peptides.

[28]  J M Thornton,et al.  LIGPLOT: a program to generate schematic diagrams of protein-ligand interactions. , 1995, Protein engineering.

[29]  Sadao Kimura,et al.  A novel potent vasoconstrictor peptide produced by vascular endothelial cells , 1988, Nature.

[30]  L. Brunton,et al.  The quasi-irreversible nature of endothelin binding and G protein-linked signaling in cardiac myocytes. , 1997, The Journal of pharmacology and experimental therapeutics.

[31]  M. Yano,et al.  [Ala1,3,11,15]endothelin-1 analogs with ETB agonistic activity. , 1991, Biochemical and biophysical research communications.

[32]  R. Stevens,et al.  Structure of an Agonist-Bound Human A2A Adenosine Receptor , 2011, Science.

[33]  Stephen M. Husbands,et al.  Structural insights into μ-opioid receptor activation , 2015, Nature.

[34]  V. Cherezov,et al.  Crystallizing membrane proteins using lipidic mesophases , 2009, Nature Protocols.

[35]  Annamaria Biroccio,et al.  β-Arrestin links endothelin A receptor to β-catenin signaling to induce ovarian cancer cell invasion and metastasis , 2009, Proceedings of the National Academy of Sciences.

[36]  J. Ballesteros,et al.  [19] Integrated methods for the construction of three-dimensional models and computational probing of structure-function relations in G protein-coupled receptors , 1995 .

[37]  Aashish Manglik,et al.  Propagation of conformational changes during μ-opioid receptor activation , 2015, Nature.

[38]  Vincent B. Chen,et al.  Correspondence e-mail: , 2000 .

[39]  Albert C. Pan,et al.  The Dynamic Process of β2-Adrenergic Receptor Activation , 2013, Cell.

[40]  D. Kohan,et al.  Regulation of blood pressure and salt homeostasis by endothelin. , 2011, Physiological reviews.

[41]  Ruben Abagyan,et al.  Crystal structure of the chemokine receptor CXCR4 in complex with a viral chemokine , 2015, Science.

[42]  M. Meens,et al.  Endothelin-1, an Endogenous Irreversible Agonist in Search of an Allosteric Inhibitor , 2009 .

[43]  Vadim Cherezov,et al.  Allosteric sodium in class A GPCR signaling. , 2014, Trends in biochemical sciences.

[44]  Yuji Kobayashi,et al.  Distributed computing and NMR constraint-based high-resolution structure determination: applied for bioactive Peptide endothelin-1 to determine C-terminal folding. , 2004, Journal of the American Chemical Society.

[45]  S. Rasmussen,et al.  Crystal Structure of the β2Adrenergic Receptor-Gs protein complex , 2011, Nature.

[46]  N. Perico,et al.  New therapeutics that antagonize endothelin: promises and frustrations , 2002, Nature Reviews Drug Discovery.

[47]  A. Davenport,et al.  Comparison of human ETA and ETB receptor signalling via G-protein and β-arrestin pathways. , 2012, Life sciences.

[48]  K. Tani,et al.  Thermostabilization of the Human Endothelin Type B Receptor. , 2016, Journal of molecular biology.

[49]  H. Kuroda,et al.  Structure-activity relationship of endothelin: importance of charged groups. , 1989, Biochemical and biophysical research communications.

[50]  W. Weis,et al.  Modified T4 Lysozyme Fusion Proteins Facilitate G Protein-Coupled Receptor Crystallogenesis. , 2014, Structure.

[51]  Macitentan: entry-into-humans study with a new endothelin receptor antagonist , 2011, European Journal of Clinical Pharmacology.

[52]  F. Spinella,et al.  Endothelin 1 in cancer: biological implications and therapeutic opportunities , 2013, Nature Reviews Cancer.

[53]  Nicholas Sawyer,et al.  Protein–protein interactions: General trends in the relationship between binding affinity and interfacial buried surface area , 2013, Protein science : a publication of the Protein Society.

[54]  Y. Hiroaki,et al.  Characterization of human endothelin B receptor and mutant receptors expressed in insect cells. , 1997, European journal of biochemistry.

[55]  R. Stevens,et al.  High-Resolution Crystal Structure of an Engineered Human β2-Adrenergic G Protein–Coupled Receptor , 2007, Science.

[56]  T. Watanabe,et al.  Characterization of endothelin receptors ETA and ETB expressed in COS cells. , 1992, Journal of biochemistry.

[57]  Eric Gouaux,et al.  A fluorescence-detection size-exclusion chromatography-based thermostability assay for membrane protein precrystallization screening. , 2012, Structure.

[58]  M. Polokoff,et al.  Endothelins: molecular biology, biochemistry, pharmacology, physiology, and pathophysiology. , 1994, Pharmacological reviews.

[59]  D. Bassolino,et al.  Conformational isomerism of endothelin in acidic aqueous media: a quantitative NOESY analysis. , 1992, Biochemistry.

[60]  B. Bremnes,et al.  Regulation and Intracellular Trafficking Pathways of the Endothelin Receptors* , 2000, The Journal of Biological Chemistry.

[61]  Gebhard F. X. Schertler,et al.  The structural basis of agonist-induced activation in constitutively active rhodopsin , 2011, Nature.

[62]  Randy J. Read,et al.  Phenix - a comprehensive python-based system for macromolecular structure solution , 2012 .