Abstract
We have studied the effects ofD-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (D-PDMP) and itsL-enantiomer on glycosphingolipids in cultured normal human kidney proximal tubular cells. We found thatD-PDMP exerted a concentration-dependent reduction in the metabolic labelling and cellular levels of glucosylceramide (GlcCer), lactosylceramide (LacCer), and the globo-series glycosphingolipids, GbOse3Cer and GbOse4Cer. It also directly inhibited the activity of UDP-glucose:ceramide β1 → 4-glucosyltransferase (GlcT-1) and UDP-galactose: GlcCer β1 → 4 galactosyltransferase (GalT-2). In contrast,L-PDMP had opposite effects on the metabolic labelling of GlcCer, LacCer, and GbOse3Cer. The levels of GlcCer and LacCer were increased, while the labelling and level of GbOse4Cer were strongly reduced. Purified GalT-2 from human kidney was inhibited byD-PDMP and stimulated byL-PDMP. It appears likely that the different glycosphingolipid glycosyltransferases possess similar binding sites for the ceramide moiety, which are blocked by binding toD-PDMP and, in the case of GbOse4Cer synthase, byL-PDMP as well. The stimulatory effects ofL-PDMP on GlcCer and LacCer synthases may be the result of binding to a modulatory site on the glycosyltransferases; in intact cells, the enzyme-analog complex may afford protection against the normal catabolic inactivation of the enzymes.
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Abbreviations
- GalT-2:
-
UDP-galactose:GlcCer β-galactosyltransferase
- GbOse3Cer:
-
Galα1 → 4Galβ1 → GlcCer
- GbOse4Cer:
-
GalNAc1 → 3Galα1 → 4Galβ1 → GlcCer
- GlcCer:
-
glucosylceramide
- GlcT-1:
-
UDP-glucose:ceramide β-glucosyltransferase
- GSLs:
-
glycosphingolipids
- LacCer:
-
lactosylceramide
- PDMP:
-
threo-1-phenyl-2-decanolyamino-3-morpholino-1-propanol
References
Chatterjee S (1991)Biochem Biophys Res Comm 181: 554–61.
Hakomori S (1990)J Biol Chem 265: 18713–16.
Obeid LM, Linardic CM, Karolak LA, Hannun YA (1993)Science 259: 1769–71.
Basu M, De T, Das KK, Kyle JW, Chon HC, Shaper RJ, Basu S (1987)Methods Enzymol 138: 575–607.
Chatterjee S, Ghosh N, Khurana, S (1992)J Biol Chem 267: 7148–53.
Shayman JA, Deshmukh GD, Mahdiyoun S, Thomas TP, Wu D, Barcelon FS, Radin NS (1991)J Biol Chem 266: 22968–74.
Marsh NL, Elias PM, Holleran WM (1995)J Clin Invest 95: 2903–9.
Datta SC, Radin NS (1988)Lipids 23: 508–10.
Shukla GS, Shukla A, Radin NS (1991)J Neurochem 56: 2125–32.
Radin NS, Shayman JA, Inokuchi J (1993) InAdvances in Lipid Research; Sphingolipids in Signaling, Part B (Bell RM, Hannun YA, Merrill AH, eds), Vol. 28, pp. 183–213, Orlando: Academic Press.
Radin NS, Shayman JA (1993) InNeuroProtocols: A Companion to Methods in Neurosciences (Fisher SK, Bleasdale JE, eds), Vol. 3, pp. 145–55. Orlando: Academic Press.
Inokuchi J, Jimbo M, Momosaki K, Shimeno II, Nagamatsu A, Radin NS (1990)Cancer Res 50: 6731–37.
Inokuchi J, Usuki S, Jimbo M (1995)J Biochem (Tokyo) 117: 766–73.
Deshmukh G, Radin NS, Gattone VH, Shayman JA (1994)J Lipid Res 35: 1611–18.
Chatterjee S, Sekerke CS, Kwiterovich Jr PO (1982)J Lipid Res 23: 513–22.
Ullman MD, McCluer RH (1977)J Lipid Res 18: 371–78.
Chatterjee S, Ghosh N, Castiglione E, Kwiterovich Jr PO (1988)J Biol Chem 263: 13017–23.
Ghosh S, Das KK, Daussin F, Basu S (1990)Ind J Biochem Biophys 27: 379–85.
Abe A, Radin NS, Shayman JA, Wotring LL, Zipkin RE, Sivakumar R, Ruggieri JM, Carson KG, Ganem B (1995)J Lipid Res 36: 611–21.
Lingwood CA (1994)Nephron 66: 21–28.
Lingwood CA (1992)Curr Opin Struct Biol 2: 693–700.
Svensson M, Lindstedt R, Radin NS, Svenborg C (1994)Infect Immunol 62: 4404–10.
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Chatterjee, S., Cleveland, T., Shi, WY. et al. Studies of the action of ceramide-like substances (d- andl-PDMP) on sphingolipid glycosyltransferases and purified lactosylceramide synthase. Glycoconjugate J 13, 481–486 (1996). https://doi.org/10.1007/BF00731481
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DOI: https://doi.org/10.1007/BF00731481