Am J Physiol Cell Physiol AJP: Regulatory, Integrative and Comparative Physiology
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Am J Physiol Cell Physiol 279: C40-C50, 2000;
0363-6143/00 $5.00
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Vol. 279, Issue 1, C40-C50, July 2000

PKA and arachidonic acid activation of human recombinant ClC-2 chloride channels

Kirti P. Tewari, Danuta H. Malinowska, Ann M. Sherry, and John Cuppoletti

Department of Molecular and Cellular Physiology, University of Cincinnati College of Medicine, Cincinnati, Ohio 45267-0576

An HEK-293 cell line stably expressing the human recombinant ClC-2 Cl- channel was used in patch-clamp studies to study its regulation. The relative permeability Px/PCl calculated from reversal potentials was I- > Cl- = NO3- = SCN->= Br-. The absolute permeability calculated from conductance ratios was Cl- = Br- = NO3- >=  SCN- > I-. The channel was activated by cAMP-dependent protein kinase (PKA), reduced extracellular pH, oleic acid (C:18 cisDelta 9), elaidic acid (C:18 transDelta 9), arachidonic acid (AA; C:20 cisDelta 5,8,11,14), and by inhibitors of AA metabolism, 5,8,11,14-eicosatetraynoic acid (ETYA; C:20 transDelta 5,8,11,14), alpha -methyl-4-(2-methylpropyl)benzeneacetic acid (ibuprofen), and 2-phenyl-1,2-benzisoselenazol-3-[2H]-one (PZ51, ebselen). ClC-2 Cl- channels were activated by a combination of forskolin plus IBMX and were inhibited by the cell-permeant myristoylated PKA inhibitor (mPKI). Channel activation by reduction of bath pH was increased by PKA and prevented by mPKI. AA activation of the ClC-2 Cl- channel was not inhibited by mPKI or staurosporine and was therefore independent of PKA or protein kinase C activation.

pH-activated ion channels; gastric HCl secretion; lung chloride channels; cystic fibrosis; nonsteroidal anti-inflammatory agents; ibuprofen; ebselen; 5,8,11,14-eicosatetraynoic acid


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