Ganglioside biosynthesis in Golgi apparatus: New perspectives on its mechanism

Yusuf, Harun K. M.; Pohlentz, Gottfried; Sandhoff, Konrad

doi:10.1002/jnr.490120205

Cited by 33 publications

(9 citation statements)

References 60 publications

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“…Of interest, the binding of the HRV strain Wa was not affected by PDMP, in agreement with the suggestion that nar3 and Wa, despite having an infectivity resistant to neuraminidase treatment of cells, bind to different cell surface sites (41). Finally, the fact that PDMP, but not tunicamycin, affected the attachment of nar3 suggests that the inhibition caused by the N-glycosylation inhibitor is not due to its reported ability to inhibit ganglioside biosynthesis (50,58).…”

Section: Discussionsupporting

confidence: 82%

Biochemical Characterization of Rotavirus Receptors in MA104 Cells

Guerrero¹,

Zárate²,

Corkidi

et al. 2000

J Virol

102

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We have tested the effect of metabolic inhibitors, membrane cholesterol depletion, and detergent extraction of cell surface molecules on the susceptibility of MA104 cells to infection by rotaviruses. Treatment of cells with tunicamycin, an inhibitor of protein N glycosylation, blocked the infectivity of the SA-dependent rotavirus RRV and its SA-independent variant nar3 by about 50%, while the inhibition of O glycosylation had no effect. The inhibitor of glycolipid biosynthesis d,l-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP) blocked the infectivity of RRV, nar3, and the human rotavirus strain Wa by about 70%. Sequestration of cholesterol from the cell membrane with ␤-cyclodextrin reduced the infectivity of the three viruses by more than 90%. The involvement of N-glycoproteins, glycolipids, and cholesterol in rotavirus infection suggests that the virus receptor(s) might be forming part of lipid microdomains in the cell membrane. MA104 cells incubated with the nonionic detergent octyl-␤-glucoside (OG) showed a ca. 60% reduction in their ability to bind rotaviruses, the same degree to which they became refractory to infection, suggesting that OG extracts the potential virus receptor(s) from the cell surface. Accordingly, when preincubated with the viruses, the OG extract inhibited the virus infectivity by more than 95%. This inhibition was abolished when the extract was treated with either proteases or heat but not when it was treated with neuraminidase, indicating the protein nature of the inhibitor. Two protein fractions of around 57 and 75 kDa were isolated from the extract, and these fractions were shown to have rotavirus-blocking activity. Also, antibodies to these fractions efficiently inhibited the infectivity of the viruses in untreated as well as in neuraminidase-treated cells. Five individual protein bands of 30, 45, 57, 75, and 110 kDa, which exhibited virus-blocking activity, were finally isolated from the OG extract. These proteins are good candidates to function as rotavirus receptors.

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Section: Discussionsupporting

confidence: 82%

Biochemical Characterization of Rotavirus Receptors in MA104 Cells

Guerrero¹,

Zárate²,

Corkidi

et al. 2000

J Virol

102

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“…Yusuf et al [57][58][59] showed that tunicamycin inhibits the synthesis of GM 1 and GM 2 gangliosides in isolated Golgi vesicles; this inhibition was found to be due to a block in carriermediated transport of nucleotide sugars across the Golgi vesicles, consistent with the luminal orientation of glycosyltransferases involved in GM2 and GMz synthesis. Yusuf et al [57][58][59] showed that tunicamycin inhibits the synthesis of GM 1 and GM 2 gangliosides in isolated Golgi vesicles; this inhibition was found to be due to a block in carriermediated transport of nucleotide sugars across the Golgi vesicles, consistent with the luminal orientation of glycosyltransferases involved in GM2 and GMz synthesis.…”

Section: Topography Of Glycolipid Glycosylation In the Golgi Apparatusmentioning

confidence: 93%

Glycolipid transfer protein and intracellular traffic of glucosylceramide

Sasaki

1990

Experientia

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Glycolipid transfer protein (GL-TP), a nonglycosylated protein with a molecular weight of 22,000 K, has been purified from pig brain. The protein transfers, by a carrier mechanism, glycolipids with a beta-glucosyl or beta-galactosyl residue directly linked to either ceramide or diacylglycerol. GL-TP appears to be present in most animal cells, and evidence has been obtained which indicates that it is a cytoplasmic protein. Little is known about the function of GL-TP. Current evidence indicates that glycosphingolipid glycosylation occurs at the luminal side of the Golgi apparatus, except for the glucosylation of ceramide, which has been shown to occur at the cytoplasmic side of the Golgi or endoplasmic membrane. It appears most likely that GL-TP participates in the intracellular traffic of glucosylceramide.

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“…Anionic membrane lipids have been identifi ed as stimulating lipid modifi ers of many membrane-associated reactions, e.g., phosphatidylglycerol (PG) stimulates steps of ganglioside biosynthesis at the Golgi compartment ( 27,28 ). BMP, phosphatidic acid, phosphatidylserine, and phosphatidylinositol (PI) stimulate the transfer of cholesterol by Niemann-Pick protein type C2 (NPC2) (29)(30)(31)(32).…”

mentioning

confidence: 99%

Membrane lipids regulate ganglioside GM2 catabolism and GM2 activator protein activity

Anheuser

Breiden

Schwarzmann

et al. 2015

Journal of Lipid Research

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For the effective degradation of lysosomal sphingolipids with short oligosaccharide head groups, special lipid binding and transfer proteins (the sphingolipid activator proteins) are needed. The sphingolipid activator proteins consist of the GM2 activator protein (GM2AP) and the saposins (Saps) A-D. They all facilitate the interaction of the membrane-bound substrate with the water-soluble enzyme at the water-membrane interface and differ from sphingolipid transfer proteins of the cytosol ( 6 ). The anionic lysosomal phospholipid, BMP, is also known to dramatically enhance (glyco)sphingolipid hydrolysis by lysosomal hydrolases and sphingolipid activator proteins ( 7-9 ).Even at low pH values (pH <5.0), BMP conveys negative surface charge to luminal lysosomal vesicles ( 10,11 ). This favors an electrostatic binding of the positively charged activator proteins and the polycationic hydrolases, and enhances degradation of (glyco)sphingolipids substantially ( 7,9,10,(12)(13)(14)(15)(16).The GM2AP, which has a hydrophobic binding cavity for lipids and a binding site for ␤ -hexosaminidase A (HexA) ( 17-19 ), arranges the interaction of ganglioside GM2 with HexA and facilitates the formation of a Michaelis-Menten complex with access of the catalytic site to the carbohydrate head group ( 20,21 ). The whole working mechanism, Abstract Ganglioside GM2 is the major lysosomal storage compound of Tay-Sachs disease. It also accumulates in Niemann-Pick disease types A and B with primary storage of SM and with cholesterol in type C. Reconstitution of GM2 catabolism with ␤ -hexosaminidase A and GM2 activator protein (GM2AP) at uncharged liposomal surfaces carrying GM2 as substrate generated only a physiologically irrelevant catabolic rate, even at pH 4.2. However, incorporation of anionic phospholipids into the GM2 carrying liposomes stimulated GM2 hydrolysis more than 10-fold, while the incorporation of plasma membrane stabilizing lipids (SM and cholesterol) generated a strong inhibition of GM2 hydrolysis, even in the presence of anionic phospholipids. Mobilization of membrane lipids by GM2AP was also inhibited in the presence of cholesterol or SM, as revealed by surface plasmon resonance studies. These lipids also reduced the interliposomal transfer rate of 2-NBD-GM1 by GM2AP, as observed in assays using Förster resonance energy transfer. Our data raise major concerns about the usage of recombinant His-tagged GM2AP compared with untagged protein. The former binds more strongly to anionic GM2-carrying liposomal surfaces, increases GM2 hydrolysis, and accelerates intermembrane transfer of 2-NBD-GM1, but does not mobilize membrane lipids. Components of eukaryotic plasma membranes reach the intraendolysosomal vesicles by endocytotic membrane fl ow ( 1 ). Those vesicles have been identifi ed as platforms for lipid and membrane degradation ( 2 ). They differ from the limiting endosomal membrane by the absence of a glycocalix, a lower cholesterol content, and the cumulative occurrence of the anionic bis(monoacylglycero)phosphate (BMP) ...

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Ganglioside biosynthesis in Golgi apparatus: New perspectives on its mechanism

Cited by 33 publications

References 60 publications

Biochemical Characterization of Rotavirus Receptors in MA104 Cells

Biochemical Characterization of Rotavirus Receptors in MA104 Cells

Glycolipid transfer protein and intracellular traffic of glucosylceramide

Membrane lipids regulate ganglioside GM2 catabolism and GM2 activator protein activity

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