Clustering and lateral concentration of raft lipids by the MAL protein.

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.

journal_name

Mol Biol Cell

authors

Magal LG,Yaffe Y,Shepshelovich J,Aranda JF,de Marco Mdel C,Gaus K,Alonso MA,Hirschberg K

doi

10.1091/mbc.e09-02-0142

subject

Has Abstract

pub_date

2009-08-01 00:00:00

pages

3751-62

issue

16

eissn

1059-1524

issn

1939-4586

pii

E09-02-0142

journal_volume

20

pub_type

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