HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 279, Issue 23, 24673-24684, June 4, 2004

This Article Full Text Full Text (PDF) All Versions of this Article: 279/23/24673    most recent M400688200v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Li, C. Articles by Naren, A. P. Search for Related Content PubMed PubMed Citation Articles by Li, C. Articles by Naren, A. P. Social Bookmarking                      What's this?

Molecular Assembly of Cystic Fibrosis Transmembrane Conductance Regulator in Plasma Membrane^*

Chunying Li, Koushik Roy, Keanna Dandridge, and Anjaparavanda P. Naren

From the Department of Physiology, University of Tennessee Health Science Center, Memphis, Tennessee 38163

Based on electrophysiological measurements, it has been argued that the active form of cystic fibrosis trans-membrane conductance regulator (CFTR) Cl^- channel is a multimer. It has also been demonstrated that this multimerization is likely due to PDZ domain-interacting partners. Here we demonstrate that although CFTR in vitro can self-associate into multimers, which depends on PDZ-based interactions, this may not be the case in cell membrane. Using chemical cross-linking, we demonstrated that CFTR exists as a higher order complex in cell membrane. However, this higher order complex is predominantly CFTR dimers, and the PDZ-interacting partners (Na⁺/H⁺ exchanger regulatory factor-1 (NHERF1) and NHERF2) constitute 2% of this complex. Interestingly solubilizing membrane expressing CFTR in detergents such as Triton X-100, Nonidet P-40, deoxycholate, and SDS tended to destabilize the CFTR dimers and dissociate them into monomeric form. The dimerization of CFTR was tightly regulated by cAMP-dependent protein kinase-dependent phosphorylation and did not depend on the active form of the channel. In addition, the dimerization was not influenced by either the PDZ motif or its interacting partners (NHERF1 and NHERF2). We also demonstrated that other signaling-related proteins such as G and syntaxin 1A can be in this higher order complex of CFTR as well. Our studies provide a deeper understanding of how the CFTR assembly takes place in native cell membrane.

Received for publication, January 21, 2004 , and in revised form, March 31, 2004.

^* This work was supported by National Institutes of Health Grant DK58545 (to A. P. N.) and American Heart Association Southeast Affiliate Grant-in-aid 0265008B (to A. P. N.). 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.

To whom correspondence should be addressed: Dept. of Physiology, University of Tennessee Health Science Center, 420 NASH, 894 Union Ave., Memphis, TN 38163. Tel.: 901-448-3137; Fax: 901-448-7126; E-mail: anaren{at}utmem.edu.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				P. M. Haggie and A. S. Verkman Monomeric CFTR in Plasma Membranes in Live Cells Revealed by Single Molecule Fluorescence Imaging J. Biol. Chem., August 29, 2008; 283(35): 23510 - 23513. [Abstract] [Full Text] [PDF]

				J. Li, P. I. Poulikakos, Z. Dai, J. R. Testa, D. J. E. Callaway, and Z. Bu Protein Kinase C Phosphorylation Disrupts Na+/H+ Exchanger Regulatory Factor 1 Autoinhibition and Promotes Cystic Fibrosis Transmembrane Conductance Regulator Macromolecular Assembly J. Biol. Chem., September 14, 2007; 282(37): 27086 - 27099. [Abstract] [Full Text] [PDF]

				F. C. Morales, Y. Takahashi, S. Momin, H. Adams, X. Chen, and M.-M. Georgescu NHERF1/EBP50 Head-to-Tail Intramolecular Interaction Masks Association with PDZ Domain Ligands Mol. Cell. Biol., April 1, 2007; 27(7): 2527 - 2537. [Abstract] [Full Text] [PDF]

				C. Mazzochi, D. J. Benos, and P. R. Smith Interaction of epithelial ion channels with the actin-based cytoskeleton Am J Physiol Renal Physiol, December 1, 2006; 291(6): F1113 - F1122. [Abstract] [Full Text] [PDF]

				G. Lamprecht and U. Seidler The emerging role of PDZ adapter proteins for regulation of intestinal ion transport Am J Physiol Gastrointest Liver Physiol, November 1, 2006; 291(5): G766 - G777. [Abstract] [Full Text] [PDF]

				I. R. Bates, B. Hebert, Y. Luo, J. Liao, A. I. Bachir, D. L. Kolin, P. W. Wiseman, and J. W. Hanrahan Membrane Lateral Diffusion and Capture of CFTR within Transient Confinement Zones Biophys. J., August 1, 2006; 91(3): 1046 - 1058. [Abstract] [Full Text] [PDF]

				D. Trompier, M. Alibert, S. Davanture, Y. Hamon, M. Pierres, and G. Chimini Transition from Dimers to Higher Oligomeric Forms Occurs during the ATPase Cycle of the ABCA1 Transporter J. Biol. Chem., July 21, 2006; 281(29): 20283 - 20290. [Abstract] [Full Text] [PDF]

				F. Brouillard, N. Bensalem, A. Hinzpeter, D. Tondelier, S. Trudel, A. D. Gruber, M. Ollero, and A. Edelman Blue Native/SDS-PAGE Analysis Reveals Reduced Expression of the mClCA3 Protein in Cystic Fibrosis Knock-out Mice Mol. Cell. Proteomics, November 1, 2005; 4(11): 1762 - 1775. [Abstract] [Full Text] [PDF]

				C. Li, K. S. Dandridge, A. Di, K. L. Marrs, E. L. Harris, K. Roy, J. S. Jackson, N. V. Makarova, Y. Fujiwara, P. L. Farrar, et al. Lysophosphatidic acid inhibits cholera toxin-induced secretory diarrhea through CFTR-dependent protein interactions J. Exp. Med., October 3, 2005; 202(7): 975 - 986. [Abstract] [Full Text] [PDF]

				M. F. Rosenberg, A. B. Kamis, L. A. Aleksandrov, R. C. Ford, and J. R. Riordan Purification and Crystallization of the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) J. Biol. Chem., September 10, 2004; 279(37): 39051 - 39057. [Abstract] [Full Text] [PDF]