title: Functional characterization of a novel CFTR mutation P67S identified in a patient with atypical cystic fibrosis.
abstract: Cystic fibrosis (CF) is caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. CFTR serves as a cAMP-stimulated chloride channel in a wide range of epithelial tissues and its dysfunction is a hallmark of CF. Over 1400 mutations in the CFTR gene are known, but functional data exist only for a minority of the mutant channels. The aim of the present study was to functionally characterize a novel CFTR mutation identified in a patient with atypical CF. Full length sequencing of the patient's CFTR gene revealed a homozygous C to T transition at nucleotide position 331 (CCT>TCT), which results in a P67S amino acid substitution. Mutant and wild-type CFTR were heterologously expressed in Xenopus laevis oocytes. CFTR whole-cell currents were studied using the two-electrode voltage-clamp technique. Channel surface expression was assessed by a chemiluminescence assay. Expression of P67S-CFTR resulted in functional CFTR chloride channels. However, the CFTR chloride conductance observed in oocytes expressing the mutant channel averaged only 24% of that in oocytes expressing wild-type CFTR. Similarly, surface expression of the mutant channel was reduced. In contrast, the mutation did not alter the anion selectivity of the channel, and Western blot analysis indicated a similar protein expression level of mutant and wild-type CFTR. Our findings indicate that the P67S mutation reduces CFTR chloride channel function by reducing channel surface expression. The mild disease phenotype of the patient indicates that the residual function of the mutant channel is sufficient to prevent the development of severe CF symptoms.
authors.affiliation: Institute of Human Genetics, University of Erlangen-Nuremberg, Germany.
authors.forename: Cornelia
authors.initials: C
authors.lastname: Kraus
authors.forename: André
authors.initials: A
authors.lastname: Reis
authors.forename: Lutz
authors.initials: L
authors.lastname: Naehrlich
authors.forename: Jörg
authors.initials: J
authors.lastname: Dötsch
authors.forename: Christoph
authors.initials: C
authors.lastname: Korbmacher
authors.forename: Robert
authors.initials: R
authors.lastname: Rauh
mesh_terms: D000200:Action Potentials; D000293:Adolescent; D019943:Amino Acid Substitution / Q000235:genetics; D001483:Base Sequence; D003550:Cystic Fibrosis / Q000201:enzymology* / Q000235:genetics*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D005260:Female; D006801:Humans; D008969:Molecular Sequence Data; D011392:Proline / Q000235:genetics* / Q000378:metabolism; D012694:Serine / Q000235:genetics* / Q000378:metabolism
chemical_list: D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D012694:Serine; D011392:Proline