Redox reagents and divalent cations alter the kinetics of cystic fibrosis transmembrane conductance regulator channel gating.
Gating of the cystic fibrosis Cl(-) channel requires hydrolysis of ATP by its nucleotide binding folds, but how this process controls the kinetics of channel gating is poorly understood. In the present work we show that the kinetics of channel gating and presumably the rate of ATP hydrolysis depends on the species of divalent cation present and the oxidation state of the protein. With Ca(2+) as the dominant divalent cation instead of Mg(2+), the open burst duration of the channel is increased approximately 20-fold, and this change is reversible upon washout of Ca(2+). In contrast, "soft" divalent cations such as Cd(2+) interact covalently with cystic fibrosis transmembrane conductance regulator (CFTR). These metals decrease both opening and closing rates of the channel, and the effects are not reversed by washout. Oxidation of CFTR channels with a variety of oxidants resulted in a similar slowing of channel gating. In contrast, reducing agents had the opposite effect, increasing both opening and closing rates of the channel. In cell-attached patches, CFTR channels exhibit both oxidized and reduced types of gating, raising the possibility that regulation of the redox state of the channel may be a physiological mode of control of CFTR channel activity.
Harrington
M A
MA
Department of Biological Sciences, Stanford University, Stanford, California 94305-5020, USA. mharring@morehouse.edu
Gunderson
K L
KL
Kopito
R R
RR
D000255:Adenosine Triphosphate / Q000378:metabolism; D002104:Cadmium / Q000494:pharmacology; D002118:Calcium / Q000494:pharmacology*; D002413:Cations, Divalent / Q000494:pharmacology*; D002460:Cell Line; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000502:physiology*; D005033:Ethylmaleimide / Q000494:pharmacology; D006801:Humans; D015640:Ion Channel Gating / Q000187:drug effects*; D007700:Kinetics; D008051:Lipid Bilayers; D008274:Magnesium / Q000494:pharmacology*; D008564:Membrane Potentials / Q000187:drug effects; D008623:Mercaptoethanol / Q000494:pharmacology; D008861:Microsomes / Q000502:physiology*; D010084:Oxidation-Reduction; D018408:Patch-Clamp Techniques; D010714:Phosphatidylethanolamines; D010766:Phosphorylation; D011196:Potassium Permanganate / Q000494:pharmacology
C505032:CFTR protein, human; D002413:Cations, Divalent; D008051:Lipid Bilayers; D010714:Phosphatidylethanolamines; D002104:Cadmium; D011196:Potassium Permanganate; C057561:1-palmitoyl-2-oleoylphosphatidylethanolamine; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D008623:Mercaptoethanol; D000255:Adenosine Triphosphate; D008274:Magnesium; D005033:Ethylmaleimide; D002118:Calcium