Glycolipid-Enriched Membrane Domains Are Assembled into Membrane Patches by Associating with the Actin Cytoskeleton

Rodgers, William; Zavzavadjian, Joelle R.

doi:10.1006/excr.2001.5253

Cited by 61 publications

(82 citation statements)

References 34 publications

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“…This extraordinary fixation of membrane microdomains within a fluid lipid matrix obviously requires some sort of stabilization, which might involve, for example, an interconnection with components of cytoskeleton. Evidence for an interaction between specific membrane domains and the cytoskeleton in animal cells has repeatedly been reported (Kusumi and Sako, 1996;Rodgers and Zavzavadjian, 2001). Actin, spectrin and vimentin have been implicated in retarding the movement of membrane proteins in various mammalian cells (Sheetz et al, 1980;Tomishige et al, 1998;Oliferenko et al, 1999;Runembert et al, 2002).…”

Section: Discussionmentioning

confidence: 85%

Visualization of Protein Compartmentation within the Plasma Membrane of Living Yeast Cells

Malínská

Malínský

Opekarová

et al. 2003

MBoC

271

319

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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. INTRODUCTIONLipid rafts (detergent-resistant membranes, detergent-insoluble glycolipid-enriched microdomains, or glycolipid-enriched membranes) are dynamic assemblies enriched in sterols and sphingolipids occurring laterally distributed in the plasma membrane of most, if not all, eukaryotes (for reviews, see Simons and Ikonen, 1997;Brown and London, 1998). In mammalian cells cholesterol, sphingomyelin, and glycosphingolipids are the basic constituents of detergentresistant membranes (Rietveld and Simons, 1998), whereas in yeast these are ergosterol, inositolphosphoceramide, and its mannosylated derivatives (Kü bler et al., 1996;Bagnat et al., 2000). Existence of similar plasma membrane domains in plant cells can also be expected, although it has not been reported so far.A distinctive feature of lipid rafts is their insolubility in mild nonionic detergents (typically Triton X-100) at 4°C (Brown and Rose, 1992;Rietveld and Simons, 1998). Consequently, they are found floating in low-density fractions of solubilizates of mild detergent-treated membranes. Rafts obtained by this procedure selectively recruit specific membrane proteins, whereas others are excluded. The recognition of selective protein retention within the detergent-resistant domains led to a concept of lateral subcompartmentation within the plasma membrane (Simons and Ikonen, 1997;Brown and London, 1998).It was proposed that rafts form a platform for lipid (Simons and Ikonen, 2000) and protein sorting and trafficking (Simons and van Meer, 1988;Galbiati et al., 2001;Ikonen, 2001) and cell signaling (Field et al., 1997;Stauffer and Meyer, 1997;Simons and Toomre, 2000;Dykstra et al., 2001). Lipid rafts provide the cells also with a mechanism for functional and spatial control of exocytosis (Chamberlain et al., 2001) and are involved in immune cell activation (for reviews, see Dykstra et al., 2001;Galbiati et al., 2001;Katagiri et al., 2001). Accumulating evidence documents that compositionally distinct lipid microdomains are assembled ...

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Section: Discussionmentioning

confidence: 85%

Visualization of Protein Compartmentation within the Plasma Membrane of Living Yeast Cells

Malínská

Malínský

Opekarová

et al. 2003

MBoC

271

319

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“…The assembly of the TCR signaling machinery is thought to be mediated by fusion of small, dynamic rafts into one stable macrodomain [64]. Translocation to the cap region and macrodomain assembly is an active process regulated by dynamic reorganization of the cytoskeleton [65][66][67]. Expression of a deletion construct that interferes with reggie oligomerization inhibits macrodomain assembly and leads to severe defects in cytoskeletal rearrangements induced by stimulation of the cell with a mitogenic lectin [M. F. Langhorst, A. Reuter, G. Luxenhofer et al, unpublished observations].…”

Section: Cellular Function Of Reggie Proteinsmentioning

confidence: 99%

Scaffolding microdomains and beyond: the function of reggie/flotillin proteins

Langhorst

Reuter

Stuermer

2005

Cell. Mol. Life Sci.

293

280

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Abstract. Reggie/flotillin proteins are considered to be components of lipid rafts and are commonly used as marker proteins for lipid microdomains. Yet almost a decade after their discovery, the function of reggies/ flotillins is still enigmatic. In this review we summarize the present state of knowledge on reggie/flotillin structure, localization and function, and discuss the role of the proteins in development and disease. Based on insights into reggie/flotillin function and by comparison with related proteins of the so-called SPFH (Stomatin/Prohibitin/ CMLS, Cell. Mol. Life Sci. 62 (2005) 2228-2240 1420-682X/05/202228-13 DOI 10.1007/s00018-005-5166-4 © Birkhäuser Verlag, Basel, 2005 Flotillin/HflK/C) protein family, including stomatin, podocin and prohibitin, we propose the existence of specific types of protein-defined microdomains which are sculpt by the clustering of individual SPFH proteins. As 'specialized rafts' similar to caveolae, these membrane domains provide platforms for the recruitment of multiprotein complexes. Since, under certain circumstances, reggie-2/flotillin-1 translocates to the nucleus, reggie/ flotillin microdomains are not only stable scaffolds but also dynamic units with their own regulatory functions.Key words. Lipid raft microdomains; signaling scaffolds; actin cytoskeleton; SPFH domain; stomatin; podocin; prohibitin; caveolae. Discovery of reggie/flotillins'Third time is a charm' the saying goes, and accordingly, reggie/flotillin proteins were independently 'discovered' three times. Madeleine Duvic and co-workers were the very first to discover part of one of the proteins during a screen for the antigen of the monoclonal antibody ECS-1 in 1994. They identified a complementary DNA (cDNA) coding for a N-terminally truncated version of reggie-1/flotillin-2 of 42 kDa (see below), which they called epidermal surface antigen (ESA) [1]. This name was later abandoned, as it was shown that this protein is not the true antigen recognized by ECS-1 [2]. In a screen for proteins upregulated in retinal ganglion cells during axon regeneration after optic nerve lesion in goldfish, we identified in 1997 two proteins of 47 kDa which we called reggie-1 and -2 [3,4]. In the same year Michael Lisanti's group identified two proteins associated with the 'floating' lipid raft fraction from mouse lung tissue which they called flotillin-1 (= reggie-2) and flotillin-2 (= reggie-1) [5]. Although flotillins became the more commonly used name for the proteins, we will stick with reggie-1 and -2, since we believe that our names and numbers are more appropriate in light of the physiological relevance of the two proteins. Structure of reggiesReggie proteins are highly conserved, with about 64% homology between fly and man [6,7]. Reggie-like proteins even exist in some bacteria, plants and fungi [8,9].

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“…Conjugates-Antibody-mediated cross-linking of certain raftassociating membrane components has been reported to induce at least a partial colocalization with other raft markers as observed by confocal microscopy in Jurkat cells (8,11,36). We accordingly used confocal imaging to compare the cell surface distributions of lipid-PEG-streptavidin conjugates with those of GM1-bound CTB and endogenous CD59 in live cells when to CTB-pretreated cells, goat anti-mouse antibody (50 g/ml) to anti-CD59-treated cells, or anti-streptavidin (SAv) antiserum (1:250) to cells pretreated with di16:0PE-PEG-biotin (5 M) followed by streptavidin (30 g/ml).…”

Section: Cell Surface Distribution Of Diacyl Pe-peg-streptavidinmentioning

confidence: 99%

Artificially Lipid-anchored Proteins Can Elicit Clustering-induced Intracellular Signaling Events in Jurkat T-Lymphocytes Independent of Lipid Raft Association

Wang¹,

Leventis

Silvius

2005

Journal of Biological Chemistry

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We have incorporated artificial lipid-anchored streptavidin conjugates with fully saturated or polyunsaturated lipid anchors into the plasma membranes of Jurkat T-lymphocytes to assess previous conclusions that the activation of signaling processes induced in these cells by clustering of endogenous glycosylphosphatidylinositol-anchored proteins or ganglioside GM1 depends specifically on the association of these membrane components with lipid rafts. Lipid-anchored streptavidin conjugates could be incorporated into Jurkat or other mammalian cell surfaces by inserting biotinylated phosphatidylethanolamine-polyethyleneglycols (PE-PEGs) and subsequently binding streptavidin to the cell-incorporated PE-PEGs. Saturated dipalmitoyl-PE-PEG-streptavidin conjugates prepared in this manner partitioned substantially into the detergent-insoluble membrane fraction isolated from Jurkat or fibroblast cells, whereas polyunsaturated dilinoleoyl-PE-PEG-anchored conjugates were wholly excluded from this fraction, consistent with the differences in the affinities of the two types of lipid anchors for liquidordered membrane domains. Remarkably, however, antibody-mediated cross-linking of either dipalmitoylor dilinoleoyl-PE-PEG-anchored streptavidin conjugates in Jurkat cells induced elevation of cytoplasmic calcium levels and tyrosine phosphorylation of the scaffolding protein linker of T-cell activation in a manner similar to that observed upon cross-linking of endogenous CD59 or ganglioside GM1. The amplitude of the cross-linking-stimulated elevation of cytoplasmic calcium moreover showed an essentially identical dependence on the level of incorporated streptavidin conjugate for either type of lipid anchor. Confocal fluorescence microscopy revealed that PE-PEG-streptavidin conjugates with saturated versus polyunsaturated anchors showed very similar surface distributions vis à vis GM1 or CD59 under conditions where one or both species were cross-linked. These results indicate that crosslinking of diverse proteins anchored only to the outer leaflet of the plasma membrane can induce activation of Jurkat T-cell-signaling responses, but they appear to contradict previous suggestions that this phenomenon rests specifically on the association of such species with lipid rafts.

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Glycolipid-Enriched Membrane Domains Are Assembled into Membrane Patches by Associating with the Actin Cytoskeleton

Cited by 61 publications

References 34 publications

Visualization of Protein Compartmentation within the Plasma Membrane of Living Yeast Cells

Visualization of Protein Compartmentation within the Plasma Membrane of Living Yeast Cells

Scaffolding microdomains and beyond: the function of reggie/flotillin proteins

Artificially Lipid-anchored Proteins Can Elicit Clustering-induced Intracellular Signaling Events in Jurkat T-Lymphocytes Independent of Lipid Raft Association

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