title: Phloxine B interacts with the cystic fibrosis transmembrane conductance regulator at multiple sites to modulate channel activity.
abstract: The fluorescein derivative phloxine B is a potent modulator of the cystic fibrosis transmembrane conductance regulator (CFTR). Low micromolar concentrations of phloxine B stimulate CFTR Cl(-) currents, whereas higher concentrations of the drug inhibit CFTR. In this study, we investigated the mechanism of action of phloxine B. Phloxine B (1 microm) stimulated wild-type CFTR and the most common cystic fibrosis mutation, DeltaF508, by increasing the open probability of phosphorylated CFTR Cl(-) channels. At each concentration of ATP tested, the drug slowed the rate of channel closure without altering the opening rate. Based on the effects of fluorescein derivatives on transport ATPases, these data suggest that phloxine B might stimulate CFTR by binding to the ATP-binding site of the second nucleotide-binding domain (NBD2) to slow the dissociation of ATP from NBD1. Channel block by phloxine B (40 microm) was voltage-dependent, enhanced when external Cl(-) concentration was reduced and unaffected by ATP (5 mm), suggesting that phloxine B inhibits CFTR by occluding the pore. We conclude that phloxine B interacts directly with CFTR at multiple sites to modulate channel activity. It or related agents might be of value in the development of new treatments for diseases caused by the malfunction of CFTR.
authors.affiliation: Department of Physiology, University of Bristol, School of Medical Sciences, University Walk, Bristol BS8, United Kingdom.
authors.forename: Zhiwei
authors.initials: Z
authors.lastname: Cai
authors.forename: David N
authors.initials: DN
authors.lastname: Sheppard
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D001665:Binding Sites; D002478:Cells, Cultured; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000737:chemistry / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004594:Electrophysiology; D010697:Eosine I Bluish / Q000737:chemistry / Q000494:pharmacology*; D005456:Fluorescent Dyes / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D051379:Mice; D008956:Models, Chemical; D009154:Mutation; D015221:Potassium Channels / Q000378:metabolism; D011485:Protein Binding; D013997:Time Factors
chemical_list: C505032:CFTR protein, human; D005456:Fluorescent Dyes; D015221:Potassium Channels; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D000255:Adenosine Triphosphate; D010697:Eosine I Bluish