1. Distribution of Can1p into stable domains reflects lateral protein segregation within the plasma membrane of living S. cerevisiae cells.
- Author
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Malinska K, Malinsky J, Opekarova M, and Tanner W
- Subjects
- Actins chemistry, Actins metabolism, Antineoplastic Agents pharmacology, Bridged Bicyclo Compounds, Heterocyclic pharmacology, Cell Wall metabolism, Chromosomes metabolism, Cloning, Molecular, Cytoskeleton metabolism, DNA metabolism, Detergents pharmacology, Genotype, Green Fluorescent Proteins metabolism, Immunoblotting, Marine Toxins pharmacology, Membrane Microdomains metabolism, Membrane Proteins chemistry, Nocodazole pharmacology, Nucleotide Transport Proteins metabolism, Octoxynol pharmacology, Plasmids metabolism, Protein Structure, Tertiary, Proton-Translocating ATPases chemistry, Saccharomyces cerevisiae Proteins chemistry, Thiazoles pharmacology, Thiazolidines, Time Factors, Amino Acid Transport Systems metabolism, Amino Acid Transport Systems, Basic metabolism, Cell Membrane metabolism, Fungal Proteins metabolism, Membrane Proteins biosynthesis, Saccharomyces cerevisiae metabolism, Saccharomyces cerevisiae Proteins biosynthesis, Saccharomyces cerevisiae Proteins metabolism
- Abstract
Recently, lipid-raft-based subdomains within the plasma membrane of living Saccharomyces cerevisiae cells were visualized using green fluorescent protein fusions, and non-overlapping subdomains containing either Pma1p or Can1p were distinguished. In this study, the long-term stability of the subdomains was investigated. Experiments with latrunculin A and nocodazole ruled out the involvement of cytoskeletal components in the stabilization of the subdomains. Also a putative role of the cell wall was excluded, because protoplasting of the cells changed neither the pattern nor the stability of the subdomains. By contrast, the expected inner dynamics of the membrane subdomains was documented by FRAP experiments. Finally, two other proteins were localized within the frame of the Can1p/Pma1p plasma-membrane partition. We show that Fur4p (another H+ symporter) and Sur7p (a protein of unknown function) occupy the Can1p subdomain.
- Published
- 2004
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