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Développement De La Technologie Des Récepteurs Couplés À Un Canal

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Développement De La Technologie Des Récepteurs Couplés À Un Canal Développement de la technologie des récepteurs couplés à un canal ionique pour des études structure-fonction des récepteurs couplés aux protéines G et du canal Kir6.2 Katarzyna Niescierowicz To cite this version: Katarzyna Niescierowicz. Développement de la technologie des récepteurs couplés à un canal ionique pour des études structure-fonction des récepteurs couplés aux protéines G et du canal Kir6.2. Sciences agricoles. Université de Grenoble, 2013. Français. NNT : 2013GRENV056. tel-01067669 HAL Id: tel-01067669 https://tel.archives-ouvertes.fr/tel-01067669 Submitted on 23 Sep 2014 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. THÈSE Pour obtenir le grade de DOCTEUR DE L’UNIVERSITÉ DE GRENOBLE Spécialité : Biologie Structurale et Nanobiologie Présentée par Katarzyna Nieścierowicz Thèse dirigée par Dr. Michel Vivaudou codirigée par Dr. Christophe Moreau préparée au sein du Laboratoire Canaux dans l'École Doctorale Chimie et Sciences de Vivant Developpement de la technologie des Récepteurs Couplés à un Canal Ionique pour des études structure-fonction des Récepteurs Couplés aux Protéines G et du canal Kir6.2. Thèse soutenue publiquement le 21 Octobre 2013 devant le jury composé de : Pr. Franck FIESCHI Président Dr. Bernard Mouillac Rapporteur Dr. Gildas LOUSSOUARN Rapporteur Dr. Pierre Charnet Membre Université Joseph Fourier / Université Pierre Mendès France / Université Stendhal / Université de Savoie / Grenoble INP Acknowledgments In the first place I would like to thank Dr. Bernard Mouillac, Dr. Gildas Loussouarn, Dr. Pierre Charnet, as well as Prof. Franck Fieschi for devoting their time in order to evaluate the work presented in the following thesis. I would like to also thank Prof. Eva-Pebay Peyroula for allowing me to perform my thesis in the Institute of Structural Biology J.P. Ebel. Big thanks to Dr. Michael Vivaudou, the leader of Channels Group and my thesis Director, for giving me a chance to experience an incredible scientific adventure here in Grenoble; as well as for his good advices and unabated enthusiasm. I would like to express my special gratitude for Dr. Christophe Moreau, Co-director of my thesis, who demonstrated an angelic patience explaining me repeatedly all the subtleties of the projects we were investigating. Working with him it was a pleasure and it tought me a lot. Moreover I would like to thank Jean whose help was priceless, including finishing of this thesis report; Maria for her willing, help, and fantastic Italian spirit; Karla for linguistic help and calm; Stephanie for positive attitude and English support for this thesis; thanks to Anne-Claire, Charlotte and Jimmy. Special thanks to Gina for her friendship and support, as well as to Anne M., Takla, Ben, Jonathan, Ali and of course Argerl for scientific (but not only) conversations. Thanks to all the people working with me during these three short years which I was lucky to spend in Grenoble, experiencing undoubtly amazing time in my life. I would especially like to thank as well my wonderful family; my parents, Ewa and Krzystof, and my gradma Albina, for their love, support and encouragement during these long years of my education; my sister Dorota which became mother of my beloved niece Kalinka, for her patience and for long hours spent on our conversations by Skype. Thanks for Marek K., Marek N. and Tomek R. for they crazy ideas and source of positive energy. Dziękuję Wam Kochani! Table of contents A List of abbreviations ............................................................................................................. 14 B PREAMBLE ......................................................................................................................... 18 I Objectives of the thesis ..................................................................................................... 20 C INTRODUCTION ................................................................................................................ 22 I Ion Channel-Coupled Receptors (ICCRs) ........................................................................ 24 I.1 Composition of ICCRs. ............................................................................................. 24 I.2 Principle of artificial-ligand gated channels. ............................................................ 24 I.3 Functionality of the coupling between fusion proteins. ............................................ 25 I.4 Applications of the ICCR technology. ...................................................................... 26 II G Protein-Coupled Receptors ........................................................................................... 26 II.1 General information. ................................................................................................. 26 II.1.1 GPCR topology. ...................................................................................................................................................... 27 II.1.2 Physiological relevance .......................................................................................................................................... 28 II.1.3 Classification of GPCRs. ........................................................................................................................................ 29 II.1.4 GPCR ligands. ......................................................................................................................................................... 33 II.1.5 Therapeutic potential of biased agonists. .............................................................................................................. 34 II.1.6 Project #1a: Pre-screening of potential biased agonists. ...................................................................................... 36 II.1.7 Models of GPCR activation ................................................................................................................................... 37 II.2 GPCRs-mediated signalling ...................................................................................... 38 II.2.1 Signaling through G Protein-Coupled Receptors.................................................................................................. 38 II.2.2 β-arrestin mediated signalling. ............................................................................................................................... 44 II.2.3 The trafficking of GPCR: from the endoplasmic reticulum to the plasma membrane ....................................... 48 II.2.4 GPCR Oligomers in signal transduction ............................................................................................................... 51 II.2.5 Cholesterol regulation of GPCRs’ function .......................................................................................................... 53 II.3 Structural coverage of the GPCR superfamily. ......................................................... 55 II.3.1 Breakthrough in GPCRs crystallographic studies. ................................................................................................ 55 II.3.2 Structural similarities and differences between GPCRs subfamilies and subtypes. ........................................... 57 II.3.3 Project #1: Functional assessment of GPCR(T4L) with the ICCR technology .................................................. 62 III ATP-sensitive potassium channel (KATP) ..................................................................... 66 III.1 The molecular composition and stoichiometry of KATP channels. ........................... 66 III.2 Physiological relevance. ........................................................................................... 67 III.3 Inwardly rectifying potassium channels: Kir ............................................................ 68 III.3.1 General characteristic of Kir channels. .................................................................................................................. 68 III.3.2 Rectification. ........................................................................................................................................................... 69 III.3.3 Structure of Kir channels. ....................................................................................................................................... 70 III.3.4 Regulation of the Kir channels. .............................................................................................................................. 74 III.4 Focus on Kir6.1 and Kir6.2 channels........................................................................ 82 III.4.1 Structural features and localization. ....................................................................................................................... 82 III.4.2 Physiological regulation of Kir6.x channels. ........................................................................................................ 82 III.5 SUR receptor, a particular member of the ABC transporters family. ......................
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