Please use this identifier to cite or link to this item: http://hdl.handle.net/10261/24790
Title: Clustering and Lateral Concentration of Raft Lipids by the MAL Protein
Authors: Goldstein Magal, Lee, Yaffe, Yakie, Shepshelovich, Jeanne, Aranda, Juan Francisco, Hirschberg, Koret, Alonso, Miguel A., Marco, María del Carmen de, Gaus, Katharina
Keywords: MAL Protein
General polarization
Fluorescent protein
Glycophosphatidylinositol
Issue Date: 15-Aug-2009
Publisher: American Society for Cell Biology
Abstract: MAL, a compact hydrophobic, four-transmembrane-domain apical protein that copurifies with detergent-resistant membranes is obligatory for the machinery that sorts glycophosphatidylinositol (GPI)-anchored proteins and others to the apical membrane in epithelia. The mechanism of MAL function in lipid-raft–mediated apical sorting is unknown. We report that MAL clusters formed by two independent procedures—spontaneous clustering of MAL tagged with the tandem dimer DiHcRED (DiHcRED-MAL) in the plasma membrane of COS7 cells and antibody-mediated cross-linking of FLAG-tagged MAL—laterally concentrate markers of sphingolipid rafts and exclude a fluorescent analogue of phosphatidylethanolamine. Site-directed mutagenesis and bimolecular fluorescence complementation analysis demonstrate that MAL forms oligomers via xx intramembrane protein–protein binding motifs. Furthermore, results from membrane modulation by using exogenously added cholesterol or ceramides support the hypothesis that MAL-mediated association with raft lipids is driven at least in part by positive hydrophobic mismatch between the lengths of the transmembrane helices of MAL and membrane lipids. These data place MAL as a key component in the organization of membrane domains that could potentially serve as membrane sorting platforms.
Description: Material suplementario: 1 archivo PDF 5 Videos
Publisher version (URL): http://dx.doi.org/10.1091/mbc.E09-02-0142
URI: http://hdl.handle.net/10261/24790
ISSN: 1059-1524
Citation: Molecular Biology of the Cell Vol. 20, Issue 16, 3751-3762 (2009)
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