NDLI: Endogenously Expressed Epithelial Sodium Channel Is Present in Lipid Rafts in A6 Cells

Content Provider	Journal of Biological Chemistry (JBC)
Author	Hill, Warren G. An, Bing Johnson, John P.
Abstract	The epithelial sodium channel (ENaC) present in the kidney collecting duct, distal colon, and the lung is responsible for salt reabsorption and whole body volume regulation. It is composed of three homologous subunits, α, β, and γ, and mutations to these subunits can lead to the salt wasting disease pseudohypoaldosteronism type I, associated with decreased channel density at the plasma membrane or to the hypertensive disorder, Liddle's syndrome, in which channel residency time at the plasma membrane is enhanced. Regulation of ENaC trafficking and turnover is therefore critical to sodium homeostasis. In this study we examined whether ENaC is present in the cholesterol-enriched microdomains commonly called lipid rafts, in the endogenously expressing A6 cell line. We demonstrate that a fraction of α, β, and γ ENaC is present in detergent-insoluble membranes, that subunits exist in membranes that float on discontinuous sucrose density gradients, and that methyl-β-cyclodextrin treatment causes a redistribution of ENaC subunits to higher density membranes. Furthermore, chronic aldosterone stimulation results in a shift in the membrane density of all three subunits. Biotinylation of apical membrane proteins revealed that ENaC is present in lipid rafts on the plasma membrane. In conclusion, these results show that ENaC is present in lipid rafts both intracellularly and on the cell surface. Raft association may be important for trafficking and/or function of the channel.
Related Links	http://www.jbc.org/content/277/37/33541.abstract
Ending Page	33544
Starting Page	33541
Page Count	4
File Format	HTM / HTML PDF
ISSN	00219258
Journal	Journal of Biological Chemistry (JBC)
Issue Number	37
Volume Number	277
DOI	10.1074/jbc.C200309200
e-ISSN	1083351X
Language	English
Publisher	American Society for Biochemistry and Molecular Biology
Publisher Date	2002-09-13
Access Restriction	Open
Subject Keyword	Epithelial sodium channel (ENaC) Protease inhibitor (PI) Post-nuclear supernatant (PNS) Phosphate-buffered saline (PBS) 4-morpholineethanesulfonic acid (MES) Methyl-β-cyclodextrin (CD) Madin-Darby canine kidney (MDCK)
Content Type	Text
Resource Type	Article
Subject	Cell Biology Biochemistry Molecular Biology

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