Visualization of Protein Compartmentation within the Plasma Membrane of Living Yeast Cells
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Published:2003-11
Issue:11
Volume:14
Page:4427-4436
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ISSN:1059-1524
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Container-title:Molecular Biology of the Cell
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language:en
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Short-container-title:MBoC
Author:
Malínská Katerina1, Malínský Jan2, Opekarová Miroslava3, Tanner Widmar1
Affiliation:
1. Universität Regensburg, Lehrstuhl für Zellbiologie und Pflanzenphysiologie, 93040 Regensburg, Germany 2. Institute of Experimental Medicine, CAS, and First Faculty of Medicine, Charles University, 12801 Prague 2, Czech Republic 3. Institute of Microbiology, Czech Academy of Science, 14220 Prague 4, Czech Republic
Abstract
Different distribution patterns of the arginine/H+symporter Can1p, the H+plasma membrane ATPase Pma1p, and the hexose transport facilitator Hxt1p within the plasma membrane of living Saccharomyces cerevisiae cells were visualized using fluorescence protein tagging of these proteins. Although Hxt1p-GFP was evenly distributed through the whole cell surface, Can1p-GFP and Pma1p-GFP were confined to characteristic subregions in the plasma membrane. Pma1p is a well-documented raft protein. Evidence is presented that Can1p, but not Hxt1p, is exclusively associated with lipid rafts, too. Double labeling experiments with Can1p-GFP– and Pma1p-RFP–containing cells demonstrate that these proteins occupy two different nonoverlapping membrane microdomains. The size of Can1p-rich (Pma1p-poor) areas was estimated to 300 nm. These domains were shown to be stable in growing cells for >30 min. To our knowledge, this is the first observation of a cell polarization-independent lateral compartmentation in the plasma membrane of a living cell.
Publisher
American Society for Cell Biology (ASCB)
Subject
Cell Biology,Molecular Biology
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