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J. Biol. Chem., Vol. 276, Issue 22, 18673-18680, June 1, 2001
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From the We have studied the regulation of
Ca2+-dependent chloride
(ClCa) channels in a human pancreatoma epithelial cell line
(CFPAC-1), which does not express functional cAMP-dependent
cystic fibrosis transmembrane conductance regulator chloride
channels. In cell-free patches from these cells, physiological
Ca2+ concentrations activated a single class of
1-picosiemens Cl
Regulation of a Human Chloride Channel
A PARADIGM FOR INTEGRATING INPUT FROM CALCIUM, TYPE II
CALMODULIN-DEPENDENT PROTEIN KINASE, AND INOSITOL
3,4,5,6-TETRAKISPHOSPHATE*
§,
, and
Inositide Signaling and Membrane
Signaling Groups, Laboratory of Signal Transduction, NIEHS, National
Institutes of Health, Research Triangle Park, North Carolina 27709 and
the ¶ Department of Molecular and Cellular Physiology, College of
Medicine, University of Cincinnati, Cincinnati, Ohio 45267-0576
-selective channels. The same channels
were also stimulated by a purified type II
calmodulin-dependent protein kinase (CaMKII), and in
cell-attached patches by purinergic agonists. In whole-cell recordings,
both Ca2+- and CaMKII-dependent mechanisms
contributed to chloride channel stimulation by Ca2+, but
the CaMKII-dependent pathway was selectively inhibited by inositol 3,4,5,6-tetrakisphosphate (Ins(3,4,5,6)P4). This
inhibitory effect of Ins(3,4,5,6)P4 on ClCa
channel stimulation by CaMKII was reduced by raising
[Ca2+] and prevented by inhibition of protein phosphatase
activity with 100 nM okadaic acid. These data provide a new
context for understanding the physiological relevance of
Ins(3,4,5,6)P4 in the longer term regulation of
Ca2+-dependent Cl fluxes in
epithelial cells.
*
This work was supported in part by National Institutes of
Health Grant DK 46433 and a grant from the Caroline Spahn Halfter Trust
Genetic Research Fund.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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