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

This Article Full Text Full Text (PDF) All Versions of this Article: 288/6/C1242    most recent 00521.2004v1 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 ISI Web of Science 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 ISI Web of Science (6) Citing Articles via Google Scholar Google Scholar Articles by Zhu, L. Articles by Forte, J. G. Search for Related Content PubMed PubMed Citation Articles by Zhu, L. Articles by Forte, J. G.

PROTEIN AND VESICLE TRAFFICKING, CYTOSKELETON

Ezrin oligomers are the membrane-bound dormant form in gastric parietal cells

¹Department of Molecular and Cell Biology, University of California, Berkeley, California; and ²Department of Pathology, University of Oklahoma Health Science Center, Oklahoma City, Oklahoma

Submitted 26 October 2004 ; accepted in final form 25 January 2005

Ezrin is a member of ezrin, radixin, moesin (ERM) protein family that links F-actin to membranes. The NH₂- and COOH-terminal association domains of ERM proteins, known respectively as N-ERMAD and C-ERMAD, participate in interactions with membrane proteins and F-actin, and intramolecular and intermolecular interactions within and among ERM proteins. In gastric parietal cells, ezrin is heavily represented on the apical membrane and is associated with cell activation. Ezrin-ezrin interactions are presumably involved in functional regulation of ezrin and thus became a subject of our study. Fluorescence resonance energy transfer (FRET) was examined with cyan fluorescent protein (CFP)- and yellow fluorescent protein (YFP)-tagged ezrin incorporated into HeLa cells and primary cultures of parietal cells. Constructs included YFP at the NH₂ terminus of ezrin (YFP-Ez), CFP at the COOH terminus of ezrin (Ez-CFP), and double-labeled ezrin (N-YFP-ezrin-CFP-C). FRET was probed using fluorescence microscopy and spectrofluorometry. Evidence of ezrin oligomer formation was found using FRET in cells coexpressing Ez-CFP and YFP-Ez and by performing coimmunoprecipitation of endogenous ezrin with fluorescent protein-tagged ezrin. Thus intermolecular NH₂- and COOH-terminal association domain (N-C) binding in vivo is consistent with the findings of earlier in vitro studies. After the ezrin oligomers were separated from monomers, FRET was observed in both forms, indicating intramolecular and intermolecular N-C binding. When the distribution of native ezrin as oligomers vs. monomers was examined in resting and maximally stimulated parietal cells, a shift of ezrin oligomers to the monomeric form was correlated with stimulation, suggesting that ezrin oligomers are the membrane-bound dormant form in gastric parietal cells.

fluorescence resonance energy transfer; acid secretion; radixin; moesin; cytoskeleton; ERM family

This article has been cited by other articles:

				L. Zhu, J. Hatakeyama, C. Chen, A. Shastri, K. Poon, and J. G. Forte Comparative study of ezrin phosphorylation among different tissues: more is good; too much is bad Am J Physiol Cell Physiol, July 1, 2008; 295(1): C192 - C202. [Abstract] [Full Text] [PDF]

				L. Zhu, R. Zhou, S. Mettler, T. Wu, A. Abbas, J. Delaney, and J. G. Forte High turnover of ezrin T567 phosphorylation: conformation, activity, and cellular function Am J Physiol Cell Physiol, September 1, 2007; 293(3): C874 - C884. [Abstract] [Full Text] [PDF]

				G. Tamma, G. Procino, M. Svelto, and G. Valenti Hypotonicity causes actin reorganization and recruitment of the actin-binding ERM protein moesin in membrane protrusions in collecting duct principal cells Am J Physiol Cell Physiol, April 1, 2007; 292(4): C1476 - C1484. [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]