Accumulation of raft lipids in T-cell plasma membrane domains engaged in TCR signalling

Zech, Tobias; Ejsing, Christer S.; Gaus, Katharina; Wet, Ben de; Shevchenko, Andrej; Simons, Kai; Harder, Thomas

doi:10.1038/emboj.2009.6

Cited by 256 publications

(243 citation statements)

References 51 publications

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“…Along the same lines, increased saturation of the hydrocarbon chains would increase interaction with Chol (35) and longer fatty acids could promote interleaflet coupling of a raft assembly (36). Consistent with earlier raft preparations, we find increased levels of 37,38). However, only isolating the apical membrane from polarized MDCK cells will tell us whether the lipids that characterize the polarized cells are indeed enriched there, however.…”

Section: Discussionsupporting

confidence: 75%

Membrane lipidome of an epithelial cell line

Sampaio

Gerl

Klose

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

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452

406

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Tissue differentiation is an important process that involves major cellular membrane remodeling. We used Madin-Darby canine kidney cells as a model for epithelium formation and investigated the remodeling of the total cell membrane lipidome during the transition from a nonpolarized morphology to an epithelial morphology and vice versa. To achieve this, we developed a shotgun-based lipidomics workflow that enabled the absolute quantification of mammalian membrane lipidomes with minimal sample processing from low sample amounts. Epithelial morphogenesis was accompanied by a major shift from sphingomyelin to glycosphingolipid, together with an increase in plasmalogen, phosphatidylethanolamine, and cholesterol content, whereas the opposite changes took place during an epithelial-to-mesenchymal transition. Moreover, during polarization, the sphingolipids became longer, more saturated, and more hydroxylated as required to generate an apical membrane domain that serves as a protective barrier for the epithelial sheet.epithelial polarity | shotgun lipidomics | high resolution mass spectrometry | Forssman glycosphingolipid | epithelial-mesenchymal transition G ain and loss of epithelial polarity are fundamental processes in tissue differentiation (1, 2). These processes have been extensively studied not only in the context of embryogenesis but in tumor progression and metastasis (1). Although cell polarization necessarily implies major changes in the structure and properties of the cell membranes, the exact changes in the molecular composition of their membrane lipidome remain unknown.Madin-Darby canine kidney (MDCK) cells are a cell culture model for epithelial polarization (3,4). When seeded at low density, MDCK cells undergo a morphological change from a fibroblast-like morphology to a well-polarized epithelium (5) and have recently also been adapted to undergo an epithelial-mesenchymal transition (EMT) (6). Epithelial cells have evolved characteristic apical and basolateral membrane domains, separated by tight junctions, with specific protein and lipid composition (7,8). The unusual robustness of the apical membrane is largely attributable to its special lipid composition (9-11).In this work, we applied shotgun MS for comprehensive and quantitative characterization of mammalian cellular membranes (12). By combining high mass resolution MS, an improved extraction procedure that increases the recovery of polar lipids (13), and the use of internal standard mixtures for absolute quantification, we expanded the lipid repertoire covered by shotgun lipidomics to glycosphingolipids (GSPs). Importantly, the analysis of the GSPs did not require the hydrolysis of glycerophospholipids (GPs) (14), and this simplification allowed us to determine the membrane lipidome with unprecedented comprehensiveness (more than 300 lipid species from 14 different lipid classes) from a single sample of 10 5 cells.We further elucidated how the lipidome is remodeled during the course of MDCK cell epithelial polarization. We found that when these c...

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Section: Discussionsupporting

confidence: 75%

Membrane lipidome of an epithelial cell line

Sampaio

Gerl

Klose

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

452

406

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“…That the T cell plasma membrane of fixed cells that are in contact with beads coated with anti--CD3 antibody is more ordered than the rest of the T cells plasma membrane, has been demonstrated by studying laurdan generalised polarisation [16,17]. In a very recent study by the same group, it was demonstrated that lipids expected to form lipid rafts like cholesterol, sphingomyelin and PC with saturated acyl chains accumulate where T cells interact with beads covered in antibodies against the T cell receptor [21]. Intriguingly, accumulation of the same lipids was seen also when T cells interacted with beads coated with antibodies against the non lipid raft marker the TfR.…”

Section: Discussionmentioning

confidence: 95%

“…Stabilisation or formation of large lipid rafts has been demonstrated to trigger all these pathways, since they can be activated by the aggregation of the ganglioside GM1 using cholera toxin B subunit (CT--B) and anti--cholera toxin [19,20]. This links reorganisation of lipids to T cell signaling, as does a recent study where it was shown that raft lipids, like cholesterol and sphingomyelin, accumulate where beads coated with TCR activating antibodies interact with a T cell [21]. However, the role of lipid rafts in TCR signalling is continuously being questioned [22--24].…”

Section: Introductionmentioning

confidence: 83%

Limited cholesterol depletion causes aggregation of plasma membrane lipid rafts inducing T cell activation

Mahammad

Dinić

Adler

et al. 2010

Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biolog

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Acute cholesterol depletion is generally associated with decreased or abolished T cell signalling but it can also cause T cell activation. This anomaly has been addressed in Jurkat T cells using progressive cholesterol depletion with methyl--beta--cyclodextrin (MBCD). At depletion levels higher than 50% there is substantial cell death, which explains reports of signalling inhibition. At 10--20% depletion levels, tyrosine phosphorylation is increased, ERK is activated and there is a small increase in cytoplasmic Ca 2+ . Peripheral actin polymerisation is also triggered by limited cholesterol depletion. Strikingly, the lipid raft marker GM1 aggregates upon cholesterol depletion and these aggregated domains concentrate the signalling proteins Lck and LAT, whereas the opposite is true for the non lipid raft marker the transferrin receptor. Using PP2, an inhibitor of Src family kinase activation, it is demonstrated that the lipid raft aggregation occurs independently of and thus upstream of the signalling response. Upon cholesterol depletion there is an increase in overall plasma membrane order, indicative of more liquid ordered domains forming at the expense of liquid disordered domains. That cholesterol depletion and not unspecific effects of MBCD was behind the reported results was confirmed by performing all experiments with MBCD--cholesterol, when no net cholesterol extraction took place. We conclude that non--lethal cholesterol depletion causes the aggregation of lipid rafts which then induces T cell signalling.

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“…Future study should focus on how the MARVEL domain-lipid interaction is used for diverse cellular tasks (Anton et al, 2008;Zech et al, 2009).…”

Section: Mechanism Of Mal and The Marvel Domainmentioning

confidence: 99%

Clustering and Lateral Concentration of Raft Lipids by the MAL Protein

Magal

Yaffe

Shepshelovich

et al. 2009

MBoC

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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.

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Accumulation of raft lipids in T-cell plasma membrane domains engaged in TCR signalling

Cited by 256 publications

References 51 publications

Membrane lipidome of an epithelial cell line

Membrane lipidome of an epithelial cell line

Limited cholesterol depletion causes aggregation of plasma membrane lipid rafts inducing T cell activation

Clustering and Lateral Concentration of Raft Lipids by the MAL Protein

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