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J. Biol. Chem., Vol. 282, Issue 50, 36481-36488, December 14, 2007

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Molecular Proximity of Cystic Fibrosis Transmembrane Conductance Regulator and Epithelial Sodium Channel Assessed by Fluorescence Resonance Energy Transfer^*

Bakhrom K. Berdiev¹, Estelle Cormet-Boyaka^¶, Albert Tousson^||, Yawar J. Qadri, Henderika M. J. Oosterveld-Hut^**, Jeong S. Hong, Patricia A. Gonzales, Cathy M. Fuller, Eric J. Sorscher, Gergely L. Lukacs^¶¶, and Dale J. Benos

From the Departments of Physiology & Biophysics, Cell Biology, ^||High Resolution Imaging Facility, and Gregory Fleming James Cystic Fibrosis Research Center, The University of Alabama at Birmingham, Birmingham, Alabama 35294-0005, the ^¶Division of Pulmonary, Critical Care, and Sleep Medicine, Davis Heart and Lung Research Institute, The Ohio State University, Columbus, Ohio 43210-1252, ^**Lambert Instruments, Turfweg 4, Leutingewolde 9313 TH, The Netherlands, the Laboratory of Kidney & Electrolyte Metabolism, NHLBI, National Institutes of Health, Bethesda, Maryland 20892-1603, and the ^¶¶Department of Laboratory Medicine and Pathobiology, Program in Cell and Lung Biology, The Hospital for Sick Children Research Institute, University of Toronto, Ontario M5G 1X8, Canada

We present the evidence for a direct physical association of cystic fibrosis transmembrane conductance regulator (CFTR) and epithelial sodium channel (ENaC), two major ion channels implicated in the pathophysiology of cystic fibrosis, a devastating inherited disease. We employed fluorescence resonance energy transfer, a distance-dependent imaging technique with capability to detect molecular complexes with near angstrom resolution, to estimate the proximity of CFTR and ENaC, an essential variable for possible physical interaction to occur. Fluorescence resonance energy transfer studies were complemented with a classic biochemical approach: coimmunoprecipitation. Our results place CFTR and ENaC within reach of each other, suggestive of a direct interaction between these two proteins.

Received for publication, September 28, 2007

^* This work was supported by National Institutes of Health Grants 2RO1-DK37206-15, P50 DK53090-05, and RO1-DK075302 CFF/CFFT. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains supplemental Figs. S1–S4.

¹ To whom correspondence should be addressed: Dept. of Cell Biology, The University of Alabama at Birmingham, 1918 University Blvd., MCLM 725, Birmingham, AL 35294-0005. Tel.: 205-934-6186; Fax: 205-934-2377; E-mail: berdiev{at}uab.edu.

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