title: Control of the CFTR channel's gates.
abstract: Unique among ABC (ATP-binding cassette) protein family members, CFTR (cystic fibrosis transmembrane conductance regulator), also termed ABCC7, encoded by the gene mutated in cystic fibrosis patients, functions as an ion channel. Opening and closing of its anion-selective pore are linked to ATP binding and hydrolysis at CFTR's two NBDs (nucleotide-binding domains), NBD1 and NBD2. Isolated NBDs of prokaryotic ABC proteins form homodimers upon binding ATP, but separate after hydrolysis of the ATP. By combining mutagenesis with single-channel recording and nucleotide photolabelling on intact CFTR molecules, we relate opening and closing of the channel gates to ATP-mediated events in the NBDs. In particular, we demonstrate that two CFTR residues, predicted to lie on opposite sides of its anticipated NBD1-NBD2 heterodimer interface, are energetically coupled when the channels open but are independent of each other in closed channels. This directly links ATP-driven tight dimerization of CFTR's cytoplasmic NBDs to opening of the ion channel in the transmembrane domains. Evolutionary conservation of the energetically coupled residues in a manner that preserves their ability to form a hydrogen bond argues that this molecular mechanism, involving dynamic restructuring of the NBD dimer interface, is shared by all members of the ABC protein superfamily.
authors.affiliation: Laboratory of Cardiac/Membrane Physiology, The Rockefeller University, New York, NY 10021, USA. paola.vergani@rockefeller.edu
authors.forename: P
authors.initials: P
authors.lastname: Vergani
authors.forename: C
authors.initials: C
authors.lastname: Basso
authors.forename: M
authors.initials: M
authors.lastname: Mense
authors.forename: A C
authors.initials: AC
authors.lastname: Nairn
authors.forename: D C
authors.initials: DC
authors.lastname: Gadsby
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D001665:Binding Sites; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000737:chemistry / Q000502:physiology*; D006801:Humans; D015640:Ion Channel Gating / Q000502:physiology*; D016296:Mutagenesis; D009711:Nucleotides / Q000378:metabolism
chemical_list: D018528:ATP-Binding Cassette Transporters; D009711:Nucleotides; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D000255:Adenosine Triphosphate