title: Contribution of a leucine residue in the first transmembrane segment to the selectivity filter region in the CFTR chloride channel.
abstract: The anion selectivity and conductance of the cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel are determined predominantly by interactions between permeant anions and the narrow region of the channel pore. This narrow region has therefore been described as functioning as the "selectivity filter" of the channel. Multiple pore-lining transmembrane segments (TMs) have previously been shown to contribute to the selectivity filter region. However, little is known about the three-dimensional organization of this region, or how multiple TMs combine to determine its functional properties. In the present study we have used patch clamp recording to identify changes in channel function associated with the formation of disulfide cross-links between cysteine residues introduced into different TMs within the selectivity filter. Cysteine introduced at position L102 in TM1 was able to form disulfide bonds with F337C and T338C in TM6, two positions that are known to play key roles in determining anion permeation properties. Consistent with this proximal arrangement of L102, F337 and T338, different mutations at L102 altered anion selectivity and conductance properties in a way that suggests that this residue plays an important role in determining selectivity filter function, albeit a much lesser role than that of F337. These results suggest an asymmetric three-dimensional arrangement of the key selectivity filter region of the pore, as well as having important implications regarding the molecular mechanism of anion permeation.
authors.affiliation: Department of Physiology & Biophysics, Dalhousie University, Halifax, Nova Scotia, Canada.
authors.forename: Alexander
authors.initials: A
authors.lastname: Negoda
authors.affiliation: Department of Physiology & Biophysics, Dalhousie University, Halifax, Nova Scotia, Canada.
authors.forename: Yassine
authors.initials: Y
authors.lastname: El Hiani
authors.affiliation: Department of Physiology & Biophysics, Dalhousie University, Halifax, Nova Scotia, Canada.
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Cowley
authors.affiliation: Department of Physiology & Biophysics, Dalhousie University, Halifax, Nova Scotia, Canada. Electronic address: paul.linsdell@dal.ca.
authors.forename: Paul
authors.initials: P
authors.lastname: Linsdell
mesh_terms: D000818:Animals; D006224:Cricetinae; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000737:chemistry* / Q000502:physiology; D004220:Disulfides / Q000737:chemistry; D004229:Dithiothreitol / Q000494:pharmacology; D007930:Leucine / Q000737:chemistry; D018408:Patch-Clamp Techniques
chemical_list: D004220:Disulfides; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D007930:Leucine; D004229:Dithiothreitol
keywords: Anion selectivity; Chloride channel; Cystic fibrosis transmembrane conductance regulator; Disulfide cross-linking; Ion channel pore; Selectivity filter