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J. Biol. Chem., Vol. 277, Issue 10, 7641-7644, March 8, 2002
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From the Department of Medicine, Division of Nephrology, The
University of Alabama at Birmingham, Birmingham, Alabama 35294
Using patch clamp techniques, we found that the
epithelial sodium channel (ENaC) activity in the apical membrane of A6
distal nephron cells showed a sudden rundown beginning at 4 min after forming the inside-out configuration. This sudden rundown was prevented
by addition of anionic phospholipids such as phosphatidylinositol 4,5-bisphosphate (PIP2), phosphatidylinositol
3,4,5-trisphosphate (PIP3), and phosphatidylserine (PS) to
the "cytoplasmic" bath. Conversely, chelation of endogenous
PIP2 with anti-PIP2 antibody, hydrolysis of
PIP2 with either exogenous phospholipase C (PLC) or
activation of endogenous PLC by extracellular ATP, or application of
the positively charged molecule, poly-L-lysine, accelerated channel rundown. However, neutral phosphatidylcholine had no
effect on ENaC activity. By two-electrode voltage clamp recordings, we demonstrated that PIP2 and PIP3 significantly
increased amiloride-sensitive current in Xenopus oocytes
injected with cRNAs of rat
ACCELERATED PUBLICATION
Anionic Phospholipids Regulate Native and Expressed Epithelial
Sodium Channel (ENaC)*
,
-, 
-, and 
-ENaC. However,
PIP2 and PIP3 did not affect surface expression of ENaC, indicating that PIP2 and PIP3 regulate
ENaC at the level of the inner plasma membrane through a mechanism that
is independent of ENaC trafficking. These data suggest that anionic
phospholipids may mediate the regulation of ENaC by PLC- or
phosphoinositide 3-kinase-coupled receptors.
*
This work was supported by a National Kidney Foundation
Young Investigator Award (to H.-P. M.).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.
To whom all correspondence should be addressed: The University of
Alabama at Birmingham, Dept. of Medicine, Division of Nephrology, 1530 Third Ave. South, Sparks Bldg. 865, Birmingham, AL 35294-0017. Tel.:
205-934-3907; Fax: 205-934-1147; E-mail:
hepingma@uab.edu.
Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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