Topology of UDP-galactose cleavage in relation to N-acetyl-lactosamine formation in Golgi vesicles. Translocation of activated galactose.
UDP-galactose appears to be produced on one side of a membrane barrier, opposite the galactosyltransferases that use it as a sugar donor. The translocation of activated galactose across membranes was studied in rat submaxillary-gland microsomal vesicles and in rat liver Golgi vesicles. When these intact vesicles containing the acceptor, N-acetylglucosamine, were incubated in the presence of UDP-galactose and two inhibitors of galactosyltransferase activity, the product, N-acetyl-lactosamine, formed within the vesicles. Thus at least the galactose moiety of UDP-galactose crossed the membranes. When intact Golgi vesicles were incubated with UDP-galactose labelled in both the uridine and the galactose moieties, labelled N-acetyllactosamine was again produced in the vesicles, but less than stoichiometric amounts of the uridine label was found there. Calculation of internal and external concentrations of UMP, a major product released from the cleaved uridine moiety, showed that the vesicles were actually enriched in UMP. When free UMP was incubated with the vesicles, this enrichment did not occur. This result was direct evidence for facilitated transport of UDP-galactose into the Golgi for use by galactosyltransferase.
D000606:Amino Sugars / Q000096:biosynthesis*; D000818:Animals; D002250:Carbon Radioisotopes; D005690:Galactose / Q000378:metabolism; D006056:Golgi Apparatus / Q000201:enzymology / Q000378:metabolism*; D066298:In Vitro Techniques; D008297:Male; D008861:Microsomes / Q000378:metabolism; D051381:Rats; D011917:Rats, Inbred Lew; D012469:Salivary Glands / Q000378:metabolism; D014316:Tritium; D014531:Uridine Diphosphate Galactose / Q000378:metabolism*; D014539:Uridine Diphosphate Sugars / Q000378:metabolism*; D014542:Uridine Monophosphate / Q000378:metabolism
D000606:Amino Sugars; D002250:Carbon Radioisotopes; D014539:Uridine Diphosphate Sugars; D014316:Tritium; D014531:Uridine Diphosphate Galactose; C000458:N-acetyllactosamine; D014542:Uridine Monophosphate; D005690:Galactose