Uridine sugar nucleotides modified in their nucleoside moieties and their effect on lipid-linked saccharide formation in vitro.
Uridine diphospho glucose (UDP-Glc) and uridine diphospho N-acetylglucosamine (UDP-GlcNAc), modified in the uridine moiety by either periodate oxidation of the ribose ring or substitution at position 5 of the uracil ring with fluorine, have been tested as potential inhibitors of glucosyl monophosphoryl dolichol (Glc-P-Dol) or N,N-diacetylchitobiosyl pyrophosphoryl dolichol [GlcNAc)2-PP-Dol) assembly in chick embryo cell membranes. The periodate oxidised sugar nucleotides inhibited glycosyl transfer from their respective natural counterparts by 50% at 230 micron periodate oxidised UDP-Glc and 70 micron periodate oxidised UDP-GlcNAc respectively. Inhibition in both cases was irreversible and addition of exogenous Dol-P stimulated only the residual non-inhibited reaction. Periodate oxidised UDP-GlcNAc preferentially inhibited the transfer of GlcNAc to GlcNac-PP-Dol. The sugar nucleotide containing 5-fluorouridine were, on the other hand, alternative substrates for Glc-P-Dol or (GlcNAc)2-PP-Dol synthesis. FUDP-Glc was a good substrate for Glc-P-Dol formation; having Km and Vmax values equal to those of UDP-Glc, whereas FUDP-GlcNAc was a less efficient substrate for the formation of (GlcNAc)2-PP-Dol; having Km and Vmax values one half and one third respectively of those of UDP-GlcNAc.
D000818:Animals; D002642:Chick Embryo; D066298:In Vitro Techniques; D007700:Kinetics; D050356:Lipid Metabolism; D009844:Oligosaccharides / Q000378:metabolism*; D011103:Polyisoprenyl Phosphate Monosaccharides / Q000096:biosynthesis; D011104:Polyisoprenyl Phosphate Oligosaccharides / Q000096:biosynthesis; D013329:Structure-Activity Relationship; D014539:Uridine Diphosphate Sugars / Q000378:metabolism / Q000494:pharmacology*
D009844:Oligosaccharides; D011103:Polyisoprenyl Phosphate Monosaccharides; D011104:Polyisoprenyl Phosphate Oligosaccharides; D014539:Uridine Diphosphate Sugars; C007292:P'-di-N-acetylchitobiosyl-P(2)-dolichylpyrophosphate; C002968:dolichol-D-glucosylmonophosphate