Alpha- and beta-xylosides alter glycolipid synthesis in human melanoma and Chinese hamster ovary cells.
beta-D-Xylosides are often used to competitively inhibit proteoglycan synthesis by serving as primers for free glycosaminoglycan (GAG) chain assembly. Quite unexpectedly, we found that when human melanoma cells and Chinese hamster ovary cells are labeled with [3H] galactose in the presence of 4-methyl umbelliferyl beta-D-xyloside (Xyl beta 4MU), a large portion of the labeled acceptor does not consist of the expected GAG chains, but of the novel GM3 ganglioside-like structure: Sia-alpha 2,3-[3H]Gal beta 1, 4Xyl beta 4MU. Moreover, formation of this derivative is associated with an inhibition of glycosphingolipid synthesis by up to 78% without affecting synthesis of other [3H]Gal-labeled glycoconjugates. Inhibition occurs rapidly and equally for all glycolipid species and is partially abrogated by brefeldin A. Inhibition requires the addition of a single galactose residue to the xyloside within the lumen of the Golgi apparatus. This addition appears to be carried out by galactosyl transferase I that normally synthesizes the core region of GAG chains. Although alpha-xyloside does not inhibit proteoglycan synthesis, it is galactosylated, but not sialylated, and is nearly as effective as a beta-xyloside at inhibiting glycolipid biosynthesis. Similar results were obtained for human macrophage U937, and differentiated or undifferentiated PC12 cells. However, in neuroblastoma cell line MR23, no low molecular weight xyloside products were made and glycolipid synthesis was not inhibited. These results suggest that some of the previously documented effects of beta-xylosides might result, in part, from their inhibition of glycolipid synthesis. The mechanism of inhibition is not a direct competition for glycolipid synthesizing enzymes; rather, it is an unexplained result of formation of Gal beta 1,4Xyl-1 (alpha or beta)4MU.
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La Jolla Cancer Research Foundation, California.
Sampath
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Varki
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D000818:Animals; D000838:Anions; D020126:Brefeldin A; D016466:CHO Cells / Q000378:metabolism*; D006224:Cricetinae; D003517:Cyclopentanes / Q000494:pharmacology; D005679:G(M3) Ganglioside / Q000378:metabolism; D005690:Galactose / Q000378:metabolism; D005966:Glucuronidase / Q000378:metabolism; D006017:Glycolipids / Q000037:antagonists & inhibitors / Q000096:biosynthesis*; D006027:Glycosides / Q000378:metabolism / Q000494:pharmacology*; D006801:Humans; D006923:Hymecromone / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism; D008545:Melanoma / Q000378:metabolism*; D009447:Neuroblastoma / Q000378:metabolism; D016716:PC12 Cells / Q000378:metabolism; D014316:Tritium; D014407:Tumor Cells, Cultured
D000838:Anions; D003517:Cyclopentanes; D005679:G(M3) Ganglioside; D006017:Glycolipids; D006027:Glycosides; C010807:xylosides; D014316:Tritium; D020126:Brefeldin A; D006923:Hymecromone; C014111:methylumbelliferyl-beta-D-xyloside; D005966:Glucuronidase; D005690:Galactose