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Potentiation of cystic fibrosis transmembrane conductance regulator (CFTR) Cl- currents by the chemical solvent tetrahydrofuran.
Mol Membr Biol 2008; 25(6-7):528-38MM

Abstract

The chemical solvent tetrahydrofuran (THF) increases short-circuit current (I(sc)) in renal epithelia endogenously expressing the cystic fibrosis transmembrane conductance regulator (CFTR). To understand how THF increases I(sc), we employed the Ussing chamber and patch-clamp techniques to study cells expressing recombinant human CFTR. THF increased I(sc) in Fischer rat thyroid (FRT) epithelia expressing wild-type CFTR with half-maximal effective concentration (K(D)) of 134 mM. This THF-induced increase in I(sc) was enhanced by forskolin (10 microM), inhibited by the PKA inhibitor H-89 (10 microM) and the thiazolidinone CFTR(inh)-172 (10 microM) and attenuated greatly in FRT epithelia expressing the cystic fibrosis mutants F508del- and G551D-CFTR. By contrast, THF (100 mM) was without effect on untransfected FRT epithelia, while other solvents failed to increase I(sc) in FRT epithelia expressing wild-type CFTR. In excised inside-out membrane patches, THF (100 mM) potentiated CFTR Cl(-) channels open in the presence of ATP (1 mM) alone by increasing the frequency of channel openings without altering their duration. However, following the phosphorylation of CFTR by PKA (75 nM), THF (100 mM) did not potentiate channel activity. Similar results were obtained with the triangle upR-S660A-CFTR Cl(-) channel that is not regulated by PKA-dependent phosphorylation and using 2'deoxy-ATP, which gates wild-type CFTR more effectively than ATP. Our data suggest that THF acts directly on CFTR to potentiate channel gating, but that its efficacy is weak and dependent on the phosphorylation status of CFTR.

Authors+Show Affiliations

Department of Physiology and Pharmacology, University of Bristol, School of Medical Sciences, Bristol, UK.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

18989824

Citation

Hughes, Lauren K., et al. "Potentiation of Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) Cl- Currents By the Chemical Solvent Tetrahydrofuran." Molecular Membrane Biology, vol. 25, no. 6-7, 2008, pp. 528-38.
Hughes LK, Ju M, Sheppard DN. Potentiation of cystic fibrosis transmembrane conductance regulator (CFTR) Cl- currents by the chemical solvent tetrahydrofuran. Mol Membr Biol. 2008;25(6-7):528-38.
Hughes, L. K., Ju, M., & Sheppard, D. N. (2008). Potentiation of cystic fibrosis transmembrane conductance regulator (CFTR) Cl- currents by the chemical solvent tetrahydrofuran. Molecular Membrane Biology, 25(6-7), pp. 528-38. doi:10.1080/09687680802487967.
Hughes LK, Ju M, Sheppard DN. Potentiation of Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) Cl- Currents By the Chemical Solvent Tetrahydrofuran. Mol Membr Biol. 2008;25(6-7):528-38. PubMed PMID: 18989824.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Potentiation of cystic fibrosis transmembrane conductance regulator (CFTR) Cl- currents by the chemical solvent tetrahydrofuran. AU - Hughes,Lauren K, AU - Ju,Min, AU - Sheppard,David N, PY - 2008/11/8/pubmed PY - 2009/1/31/medline PY - 2008/11/8/entrez SP - 528 EP - 38 JF - Molecular membrane biology JO - Mol. Membr. Biol. VL - 25 IS - 6-7 N2 - The chemical solvent tetrahydrofuran (THF) increases short-circuit current (I(sc)) in renal epithelia endogenously expressing the cystic fibrosis transmembrane conductance regulator (CFTR). To understand how THF increases I(sc), we employed the Ussing chamber and patch-clamp techniques to study cells expressing recombinant human CFTR. THF increased I(sc) in Fischer rat thyroid (FRT) epithelia expressing wild-type CFTR with half-maximal effective concentration (K(D)) of 134 mM. This THF-induced increase in I(sc) was enhanced by forskolin (10 microM), inhibited by the PKA inhibitor H-89 (10 microM) and the thiazolidinone CFTR(inh)-172 (10 microM) and attenuated greatly in FRT epithelia expressing the cystic fibrosis mutants F508del- and G551D-CFTR. By contrast, THF (100 mM) was without effect on untransfected FRT epithelia, while other solvents failed to increase I(sc) in FRT epithelia expressing wild-type CFTR. In excised inside-out membrane patches, THF (100 mM) potentiated CFTR Cl(-) channels open in the presence of ATP (1 mM) alone by increasing the frequency of channel openings without altering their duration. However, following the phosphorylation of CFTR by PKA (75 nM), THF (100 mM) did not potentiate channel activity. Similar results were obtained with the triangle upR-S660A-CFTR Cl(-) channel that is not regulated by PKA-dependent phosphorylation and using 2'deoxy-ATP, which gates wild-type CFTR more effectively than ATP. Our data suggest that THF acts directly on CFTR to potentiate channel gating, but that its efficacy is weak and dependent on the phosphorylation status of CFTR. SN - 1464-5203 UR - https://www.unboundmedicine.com/medline/citation/18989824/Potentiation_of_cystic_fibrosis_transmembrane_conductance_regulator__CFTR__Cl__currents_by_the_chemical_solvent_tetrahydrofuran_ L2 - http://www.tandfonline.com/doi/full/10.1080/09687680802487967 DB - PRIME DP - Unbound Medicine ER -