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Anionic phospholipids differentially regulate the epithelial sodium channel (ENaC) by interacting with α, β, and γ ENaC subunits

  • Ion Channels, Receptors and Transporters
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Abstract

Anionic phospholipids (APs) present a variety of lipids in the cytoplasmic leaflet of the plasma membrane, including phosphatidylinositol (PI), PI-4-phosphate (PI(4)P), phosphatidylserine (PS), PI-4,5-bisphosphate (PI(4,5)P2), PI-3,4,5-trisphosphate (PI(3,4,5)P3), and phosphatidic acid (PA). We previously showed that PI(4,5)P2 and PI(3,4,5)P3 upregulate the renal epithelial sodium channel (ENaC). Further studies from others suggested that PI(4,5)P2 and PI(3,4,5)P3 respectively target β- and γ-ENaC subunit. To determine whether PI(4,5)P2 and PI(3,4,5)P3 selectively bind to β and γ subunit, we performed lipid-protein overlay experiments. Surprisingly, the results reveal that most APs, including PI(4)P, PS, PI(4,5)P2, PI(3,4,5)P3, and PA, but not PI, non-selectively bind to not only β and γ but also α subunit. To determine how these APs regulate ENaC, we performed inside-out patch-clamp experiments and found that PS, but not PI or PI(4)P, maintained ENaC activity, that PI(4,5)P2 and PI(3,4,5)P3 stimulated ENaC, and that PA, however, inhibited ENaC. These data together suggest that APs differentially regulate ENaC by physically interacting with α-, β-, and γ-ENaC. Further, the data from cell-attached patch-clamp and confocal microscopy experiments indicate that PA, a product of phospholipase D, may provide one of the pathways for inhibition of ENaC by endothelin receptors.

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Acknowledgments

We thank Albert Tousson in the Imaging Facilities at the University of Alabama at Birmingham for his technical assistance on confocal microscopy experiments.

Grants

This research was supported by the Natural Science Foundation of the People’s Republic of China (30871007 to Z-R Zhang), Natural Science Foundation of Heilongjiang Province (ZD200807-01 and ZD2008-08 to Z-R Zhang), Grant from Educational Office of Heilongjiang Province (1154HZ11 to Z-R Zhang), Department of Health and Human Services (National Institutes of Health Grant R01-DK067110 to H-P Ma), and American Society of Nephrology (M. James Scherbenske Grant to H-P Ma).

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Authors and Affiliations

  1. Departments of Pharmacy and Cardiology, The 2nd Affiliated Hospital, Key Laboratories of Education Ministry for Myocardial Ischemia Mechanism and Treatment, Harbin Medical University, Harbin, Heilongjiang Province, 150086, People’s Republic of China

    Zhi-Ren Zhang

  2. Department of Medicine, Division of Nephrology, University of Alabama, 1530 Third Avenue South, Birmingham, AL, 35294, USA

    Chu-Fang Chou, Jing Wang, You-You Liang & He-Ping Ma

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  1. Zhi-Ren Zhang
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  2. Chu-Fang Chou
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  3. Jing Wang
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  4. You-You Liang
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  5. He-Ping Ma
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Correspondence to Zhi-Ren Zhang or He-Ping Ma.

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Zhi-Ren Zhang, Chu-Fang Chou, and Jing Wang contribute equally to this work.

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Zhang, ZR., Chou, CF., Wang, J. et al. Anionic phospholipids differentially regulate the epithelial sodium channel (ENaC) by interacting with α, β, and γ ENaC subunits. Pflugers Arch - Eur J Physiol 459, 377–387 (2010). https://doi.org/10.1007/s00424-009-0733-4

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