title: Involvement of the heterodimeric interface region of the nucleotide binding domain-2 (NBD2) in the CFTR quaternary structure and membrane stability.
abstract: The cystic fibrosis transmembrane conductance regulator (CFTR) is the only member of the ATP-binding cassette (ABC) superfamily that functions as a chloride channel. The predicted structure of CFTR protein contains two membrane-spanning domains (MSDs), each followed by a nucleotide binding domain (NBD1 and NBD2). The opening of the Cl- channel is directly linked to ATP-driven tight dimerization of CFTR's NBD1 and NBD2 domains. The presence of a heterodimeric interfaces (HI) region in NBD1 and NBD2 generated a head to tail orientation necessary for channel activity. This process was also suggested to promote important conformational changes in the associated transmembrane domains of CFTR, which may impact the CFTR plasma membrane stability. To better understand the role of the individual HI region in this process, we generated recombinant CFTR protein with suppressed HI-NBD1 and HI-NBD2. Our results indicate that HI-NBD2 deletion leads to the loss of the dimerization profile of CFTR that affect its plasma membrane stability. We conclude that, in addition to its role in Cl- transport, HI-NBD2 domain confers membrane stability of CFTR by consolidating its quaternary structure through interactions with HI-NBD1 region.
authors.affiliation: Centre National de la Recherche Scientifique (CNRS), LCBM-UMR 5249, Grenoble, France; Commissariat à l'Energie Atomique (CEA), DSV-iRTSV, Grenoble, France; Grenoble Alpes Université (GAU), Grenoble 1, France.
authors.forename: Julien
authors.initials: J
authors.lastname: Micoud
authors.affiliation: Centre National de la Recherche Scientifique (CNRS), LCBM-UMR 5249, Grenoble, France; Commissariat à l'Energie Atomique (CEA), DSV-iRTSV, Grenoble, France; Grenoble Alpes Université (GAU), Grenoble 1, France.
authors.forename: Sylvain
authors.initials: S
authors.lastname: Chauvet
authors.affiliation: Laboratory of Clinical Investigation III, Geneva University Hospitals and University of Geneva, 1211 Geneva, Switzerland.
authors.forename: Klaus Ernst Ludwig
authors.initials: KE
authors.lastname: Scheckenbach
authors.affiliation: Commissariat à l'Energie Atomique (CEA), DSV-iRTSV, Grenoble, France; Grenoble Alpes Université (GAU), Grenoble 1, France; Institut National de la Santé et de la Recherche Médicale (INSERM), U1036 Grenoble, France.
authors.forename: Nadia
authors.initials: N
authors.lastname: Alfaidy
authors.affiliation: Laboratory of Clinical Investigation III, Geneva University Hospitals and University of Geneva, 1211 Geneva, Switzerland.
authors.forename: Marc
authors.initials: M
authors.lastname: Chanson
authors.affiliation: Centre National de la Recherche Scientifique (CNRS), LCBM-UMR 5249, Grenoble, France; Commissariat à l'Energie Atomique (CEA), DSV-iRTSV, Grenoble, France; Grenoble Alpes Université (GAU), Grenoble 1, France. Electronic address: mohamed.benharouga@cea.fr.
authors.forename: Mohamed
authors.initials: M
authors.lastname: Benharouga
mesh_terms: D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006224:Cricetinae; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D055550:Protein Stability; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary
chemical_list: C505032:CFTR protein, human; D019005:Cystic Fibrosis Transmembrane Conductance Regulator
keywords: CFTR; Cystic fibrosis; Dimerization; Membrane stability; NBD2; Quaternary structure