title: Defective function of the cystic fibrosis-causing missense mutation G551D is recovered by genistein.
abstract: The patch-clamp technique was used to investigate the effects of the isoflavone genistein on disease-causing mutations (G551D and DeltaF508) of the cystic fibrosis transmembrane conductance regulator (CFTR). In HeLa cells recombinantly expressing the trafficking-competent G551D-CFTR, the forskolin-stimulated Cl currents were small, and average open probability of G551D-CFTR was P(o) = 0.047 +/- 0.019. Addition of genistein activated Cl currents approximately 10-fold, and the P(o) of G551D-CFTR increased to 0.49 +/- 0.12, which is a P(o) similar to wild-type CFTR. In cystic fibrosis (CF) epithelial cells homozygous for the trafficking-impaired DeltaF508 mutation, forskolin and genistein activated Cl currents only after 4-phenylbutyrate treatment. These data suggested that genistein activated CFTR mutants that were present in the cell membrane. Therefore, we tested the effects of genistein in CF patients with the G551D mutation in nasal potential difference (PD) measurements in vivo. The perfusion of the nasal mucosa of G551D CF patients with isoproterenol had no effect; however, genistein stimulated Cl-dependent nasal PD by, on average, -2.4 +/- 0.6 mV, which corresponds to 16.9% of the responses (to beta-adrenergic stimulation) found in healthy subjects.
authors.affiliation: Research Institute and Pulmonary Center, Children's Hospital Oakland, Oakland 94609, California, USa.
authors.forename: B
authors.initials: B
authors.lastname: Illek
authors.forename: L
authors.initials: L
authors.lastname: Zhang
authors.forename: N C
authors.initials: NC
authors.lastname: Lewis
authors.forename: R B
authors.initials: RB
authors.lastname: Moss
authors.forename: J Y
authors.initials: JY
authors.lastname: Dong
authors.forename: H
authors.initials: H
authors.lastname: Fischer
mesh_terms: D001692:Biological Transport / Q000187:drug effects; D002675:Child, Preschool; D002712:Chlorides / Q000378:metabolism; D003550:Cystic Fibrosis / Q000235:genetics*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics* / Q000378:metabolism; D019833:Genistein / Q000494:pharmacology*; D006367:HeLa Cells / Q000378:metabolism / Q000502:physiology; D006801:Humans; D020125:Mutation, Missense / Q000502:physiology*; D018408:Patch-Clamp Techniques
chemical_list: C505032:CFTR protein, human; D002712:Chlorides; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D019833:Genistein