Raft-based sphingomyelin interactions revealed by new fluorescent sphingomyelin analogs

Kinoshita, Masanao; Suzuki, Kenichi; Matsumori, Nobuaki; Takada, Misa; Ano, Hikaru; Morigaki, Kenichi; Abe, Mitsuhiro; Makino, Asami; Kobayashi, Toshihide; Hirosawa, K; Fujiwara, Takeshi; Kusumi, Akihiro; Murata, Michio

doi:10.1083/jcb.201607086

Cited by 112 publications

(146 citation statements)

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“…Importantly, the recruitment of cytoplasmic signaling molecules at the CD59 signaling rafts occurred transiently, in a time scale on the order of fractions of a second (in the following text, we use the expression “recruitment of signaling molecules ‘at’ CD59 clusters” rather than “the recruitment ‘to’ CD59 clusters” because our imaging method could not directly show the binding of the signaling molecules located in the inner leaflet to the CD59 clusters located in the outer leaflet). Raftlike properties of the artificial antibody (Ab)-induced CD59 clusters were confirmed by the finding that fluorescently labeled gangliosides and sphingomyelins are colocalized with the artificial CD59 clusters ( Komura et al, 2016 ; Kinoshita et al, 2017 ). CD59-TM, in which the GPI anchor was replaced by the TM domain of a prototypical nonraft molecule, low-density lipoprotein receptor (LDLR), failed to exhibit the raftlike behaviors and to trigger the downstream signal, in ways similar to the CD59 clusters after cholesterol depletion ( Suzuki et al, 2007a , 2007b , 2012 ).…”

Section: Introductionmentioning

confidence: 92%

High-speed single-molecule imaging reveals signal transduction by induced transbilayer raft phases

Koyama-Honda

Fujiwara

Kasai

et al. 2020

Journal of Cell Biology

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Using single-molecule imaging with enhanced time resolutions down to 5 ms, we found that CD59 cluster rafts and GM1 cluster rafts were stably induced in the outer leaflet of the plasma membrane (PM), which triggered the activation of Lyn, H-Ras, and ERK and continually recruited Lyn and H-Ras right beneath them in the inner leaflet with dwell lifetimes <0.1 s. The detection was possible due to the enhanced time resolutions employed here. The recruitment depended on the PM cholesterol and saturated alkyl chains of Lyn and H-Ras, whereas it was blocked by the nonraftophilic transmembrane protein moiety and unsaturated alkyl chains linked to the inner-leaflet molecules. Because GM1 cluster rafts recruited Lyn and H-Ras as efficiently as CD59 cluster rafts, and because the protein moieties of Lyn and H-Ras were not required for the recruitment, we conclude that the transbilayer raft phases induced by the outer-leaflet stabilized rafts recruit lipid-anchored signaling molecules by lateral raft–lipid interactions and thus serve as a key signal transduction platform.

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Section: Introductionmentioning

confidence: 92%

High-speed single-molecule imaging reveals signal transduction by induced transbilayer raft phases

Koyama-Honda

Fujiwara

Kasai

et al. 2020

Journal of Cell Biology

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“…They are thought to co-exist in the plasma membrane with liquid disordered (Ld) domains, which are rich in lipids that pack loosely because they have unsaturated acyl chains (Brown and London, 1997;Schroeder et al, 1994;Veatch and Keller, 2005). Although the existence of lipid rafts in the cell plasma membrane is still not universally accepted, very recent studies have provided strong evidence for their presence in mammalian cells (Kinoshita et al, 2017;Komura et al, 2016;Stone et al, 2017). Rafts have been proposed to be involved in many biological processes, including cellular viability, differentiation, infection, signaling pathways, and the sorting of membrane lipids and proteins (Bang et al, 2005;George et al, 2012;Gniadecki, 2004;Head et al, 2014;Xu et al, 2009).…”

Section: Introductionmentioning

confidence: 99%

The effect of sterol structure upon clathrin-mediated and clathrin-independent endocytosis

Kim

Singh

Poeta

et al. 2017

Journal of Cell Science

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Ordered lipid domains (rafts) in plasma membranes have been hypothesized to participate in endocytosis based on inhibition of endocytosis by removal or sequestration of cholesterol. To more carefully investigate the role of the sterol in endocytosis, we used a substitution strategy to replace cholesterol with sterols that show various raft-forming abilities and chemical structures. Both clathrin-mediated endocytosis of transferrin and clathrin-independent endocytosis of clustered placental alkaline phosphatase were measured. A subset of sterols reversibly inhibited both clathrin-dependent and clathrin-independent endocytosis. The ability of a sterol to support lipid raft formation was necessary for endocytosis. However, it was not sufficient, because a sterol lacking a 3β-OH group did not support endocytosis even though it had the ability to support ordered domain formation. Double bonds in the sterol rings and an aliphatic tail structure identical to that of cholesterol were neither necessary nor sufficient to support endocytosis. This study shows that substitution using a large number of sterols can define the role of sterol structure in cellular functions. Hypotheses for how sterol structure can similarly alter clathrin-dependent and clathrin-independent endocytosis are discussed.

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“…GPI-APs were proposed to associate with “lipid rafts”, microdomains enriched in cholesterol and sphingolipids, from the trans-Golgi network (TGN) to the plasma membrane 18–21 . Evidence suggests that lipid rafts are highly dynamic and heterogeneous 22 , and GPI-APs are thought to be organized in cholesterol- and sphingomyelin-dependent submicron-sized domains in live cell membranes 23–26 . The current view of the functional microdomains is that they might be present in the Golgi 27–29 .…”

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confidence: 99%

Cross-talks of glycosylphosphatidylinositol biosynthesis with glycosphingolipid biosynthesis and ER-associated degradation

Wang

Maeda

Liu

et al. 2019

Preprint

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16 17 18 Glycosylphosphatidylinositol (GPI)-anchored proteins and glycosphingolipids interact with 19 each other in the mammalian plasma membranes, forming dynamic microdomains. How their 20interaction starts in the cells has been unclear. Here, based on a genome-wide CRISPR-Cas9 21 genetic screen for genes required for GPI side-chain modification by galactose in the Golgi 22 apparatus, we report that b1,3-galactosyltransferase 4 (B3GALT4), also called GM1 23 ganglioside synthase, additionally functions in transferring galactose to the N-24 acetylgalactosamine side-chain of GPI. Furthermore, B3GALT4 requires lactosylceramide 25 for the efficient GPI side-chain galactosylation. Thus, our work demonstrates previously 26 unexpected evolutionary and functional relationships between GPI-anchored proteins and 27 glycosphingolipids in the Golgi. Through the same screening, we also show that GPI 28 biosynthesis in the endoplasmic reticulum (ER) is severely suppressed by ER-associated 29 degradation to prevent GPI accumulation when the transfer of synthesized GPI to proteins is 30 defective. Our data demonstrates cross-talks of GPI biosynthesis with glycosphingolipid 31 biosynthesis and the ER quality control system. 32 33CRISPR genome-wide screen. 36 37 Glycosylphosphatidylinositol (GPI) is a complex glycolipid for post-translational 38 modification of many cell-surface proteins in eukaryotic cells 1 . To date, more than 150 39 human proteins have been confirmed as GPI-anchored proteins (GPI-APs) 2 . The structure of 40 the core backbone of GPI, which is conserved in eukaryotic organisms, is EtNP-6Mana-41 2Mana-6Mana-4GlcNa-6Inositol-phospholipid (where EtNP, Man and GlcN are 42 ethanolamine phosphate, mannose and glucosamine, respectively) ( Fig. 1a). GPI is 43 synthesized in the endoplasmic reticulum (ER) followed by transfer of GPI to proteins that 44 have a C-terminal GPI-attachment signal peptide. The GPI-attachment signal peptide is 45 removed and replaced with GPI by the GPI-transamidase (GPI-Tase) complex to form 46 immature GPI-APs. Nascent GPI-APs undergo structural remodeling in the ER and the Golgi 47 apparatus. The inositol-linked acyl chain is deacylated and the EtNP side branch is removed 48 from the second Man (Man2) for efficient ER to Golgi transport 3-5 . In the Golgi, GPI fatty 49 acid remodeling occurs, in which an sn-2-linked unsaturated fatty acyl chain is removed and 50 reacylated with a saturated chain, usually stearic acid 6,7 . Fatty acid remodeling is crucial for 51 lipid-raft association of GPI, a feature of GPI-APs 8 . 52 53The structural variation of GPI anchors is introduced by side-chain modifications 1 . Structural 54 studies of GPIs from some mammalian GPI-APs indicated that the first mannose (Man1) is 55 often modified with β1,4-linked N-acetylgalactosamine (GalNAc) 9,10 . This modification is 56 mediated by PGAP4 (Post-GPI attachment to proteins 4 also known as TMEM246), a 57 recently identified Golgi-resident, GPI-specific GalNAc-transferase 11 . The GalNAc side-58 chain can be furth...

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Raft-based sphingomyelin interactions revealed by new fluorescent sphingomyelin analogs

Cited by 112 publications

References 65 publications

High-speed single-molecule imaging reveals signal transduction by induced transbilayer raft phases

High-speed single-molecule imaging reveals signal transduction by induced transbilayer raft phases

The effect of sterol structure upon clathrin-mediated and clathrin-independent endocytosis

Cross-talks of glycosylphosphatidylinositol biosynthesis with glycosphingolipid biosynthesis and ER-associated degradation

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