Visualization of protein compartmentation within the plasma membrane of living yeast cells.
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.
Malínská
Katerina
K
Universität Regensburg, Lehrstuhl für Zellbiologie und Pflanzenphysiologie, 93040 Regensburg, Germany.
Malínský
Jan
J
Opekarová
Miroslava
M
Tanner
Widmar
W
D026905:Amino Acid Transport Systems / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000378:metabolism; D005656:Fungal Proteins / Q000378:metabolism*; D051246:Glucose Transport Proteins, Facilitative; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D021962:Membrane Microdomains / Q000378:metabolism*; D009004:Monosaccharide Transport Proteins / Q000378:metabolism*; D006180:Proton-Translocating ATPases / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*
D026905:Amino Acid Transport Systems; C097640:CAN1 protein, Candida albicans; D005656:Fungal Proteins; D051246:Glucose Transport Proteins, Facilitative; C090594:HXT1 protein, S cerevisiae; D008164:Luminescent Proteins; D009004:Monosaccharide Transport Proteins; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D049452:Green Fluorescent Proteins; C092477:PMA1 protein, S cerevisiae; D006180:Proton-Translocating ATPases