Endogenous glycosphingolipids move to the cell surface at a rate consistent with bulk flow estimates.

Young, William W.; Lutz, Mallory S.; Blackburn, W A

doi:10.1016/s0021-9258(19)49798-6

Cited by 40 publications

(3 citation statements)

References 36 publications

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“…SM (16,21,22), lactosylceramide (see reference 26), and SGalCer (50) are synthesized in the lumen of the Golgi and are unable to translocate across the Golgi membrane. Thus, after synthesis they are expected to reach the cell surface by vesicular transport, which has been experimentally confirmed for gangliosides (32,59). It is less clear how GlcCer and GalCer are transported to the cell surface.…”

Section: Topology Of Sphingolipid Synthesis and Mechanism Of Transportmentioning

confidence: 91%

Sorting of newly synthesized galactosphingolipids to the two surface domains of epithelial cells.

Bijl

Lopes‐Cardozo

Meer

1996

The Journal of Cell Biology

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Abstract. The high concentration of glycosphingolipids on the apical surface of epithelial cells may be generated by selective transport from their site of synthesis to the cell surface. Previously, we showed that canine kidney MDCK and human intestinal Caco-2 cells convetted a ceramide carrying the short fluorescent fatty acid Cr-NBD to glucosylceramide (GlcCer) and sphingomyelin (SM), and that GlcCer was preferentially transported to the apical surface as compared to SM. Here, we address the point that not all glycosphingolipid classes are apically enriched in epithelia. We show that a ceramide containing the 2-hydroxy fatty acid C60H was preferentially converted by MDCK and Caco-2 cells to galactosylceramide (GalCer) and its derivatives galabiosylceramide (Ga2Cer) and sulfatide (SGalCer) as compared to SM and GlcCer--all endogenous lipid classes of these cells. Transport to the apical and basolateral cell surface was monitored by a BSAdepletion assay. In MDCK cells, GalCer reached the cell surface with a two-to sixfold lower apical/basolateral polarity than GlcCer. Remarkably, in Caco-2 cells GalCer and GlcCer displayed the same apical/basolateral polarity, but it was sixfold lower for lipids with aCrOH chain than for C6-NBD lipids. Therefore, the sorting of a sphingolipid appears to depend on lipid structure and cell type. We propose that the different ratios of gluco-and galactosphingolipid synthesis in the various epithelial tissues govern lipid sorting in the membrane of the trans Golgi network by dictating the composition of the domains from where vesicles bud to the apical and basolateral cell surface.

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Section: Topology Of Sphingolipid Synthesis and Mechanism Of Transportmentioning

confidence: 91%

Sorting of newly synthesized galactosphingolipids to the two surface domains of epithelial cells.

Bijl

Lopes‐Cardozo

Meer

1996

The Journal of Cell Biology

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“…Therefore, we have searched for a suitable chemical procedure. Previously, chemical modification of glycolipids has been performed by a procedure employing sodium periodate (11,12) as reviewed by Sillence, Raggers, and van Meer (13). Periodate is suitable for the oxidation of biomolecules that contain vicinal hydroxyl groups.…”

mentioning

confidence: 99%

Assay for the transbilayer distribution of glycolipids: selective oxidation of glucosylceramide to glucuronylceramide by TEMPO nitroxyl radicals

Sillence

Raggers

Neville

et al. 2000

Journal of Lipid Research

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In the present study, 2,2,6,6-tetramethylpiperidinooxy nitroxide (TEMPO) has been applied successfully to discriminate between glucosylceramide in the outer and inner leaflets of closed membrane bilayers. The nitroxyl radicals TEMPO and carboxy-TEMPO, once oxidized to nitrosonium ions, are capable of oxidizing residues that contain primary hydroxyl and amino groups. When applied to radiolabeled glucosylceramide in liposomes, oxidation with TEMPO led to an oxidized product that was easily separated from the original lipid by thin-layer chromatography, and that was identified by mass spectrometric analysis as the corresponding acid glucuronylceramide. To test whether oxidation was confined to the external leaflet, TEMPO was applied to large unilamellar vesicles (LUVs) consisting of egg phosphatidylcholine-egg phosphatidylethanolamine-cholesterol 55:5:40 (mol/mol). TEMPO oxidized most radiolabeled phosphatidylethanolamine, whereas carboxy-TEMPO oxidized only half. Hydrolysis by phospholipase A 2 confirmed that 50% of the phosphatidylethanolamine was accessible in the external bilayer leaflet, suggesting that TEMPO penetrated the lipid bilayer and carboxy-TEMPO did not. When applied to LUVs containing Ͻ 1 mol% radiolabeled glucosylceramide or short-chain C 6 -glucosylceramide, carboxy-TEMPO oxidized half the glucosylceramide. However, if surface C 6 -glucosylceramide was first depleted by bovine serum albumin (BSA) (extracting 49 ؎ 1%), 94% of the remaining C 6 -glucosylceramide was resistant to oxidation. Carboxy-TEMPO oxidized glucosylceramide on the surface of LUVs without affecting inner leaflet glucosylceramide. At pH 9.5 and at 0 ؇ C, the reaction reached completion by 20 min.

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“…The sphingosine base varies in structure only to a small degree, but the acyl chain can display considerable heterogeneity in both length (C14–C26) and saturation (single cis or no double bond) ( Hannun and Obeid, 2018 ; Merrill, 2011 ). Because these lipids contain large hydrophilic extracellular head groups, they are trapped in the outer leaflet of the membrane bilayer, and their distribution throughout the cell occurs only by vesicular traffic ( van Meer et al, 2008 ; Young et al, 1992 ). This can be quantitatively measured and compared for GM1 species with identical head groups but with varied acyl chains in their ceramide moieties.…”

Section: Introductionmentioning

confidence: 99%

Structural basis for acyl chain control over glycosphingolipid sorting and vesicular trafficking

et al. 2022

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Endogenous glycosphingolipids move to the cell surface at a rate consistent with bulk flow estimates.

Cited by 40 publications

References 36 publications

Sorting of newly synthesized galactosphingolipids to the two surface domains of epithelial cells.

Sorting of newly synthesized galactosphingolipids to the two surface domains of epithelial cells.

Assay for the transbilayer distribution of glycolipids: selective oxidation of glucosylceramide to glucuronylceramide by TEMPO nitroxyl radicals

Structural basis for acyl chain control over glycosphingolipid sorting and vesicular trafficking

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