Glycolipid intermediates involved in the transfer of N-acetylglucosamine to endogenous proteins in a yeast membrane preparation.
A particulate membrane fraction from Saccharomyces cerevisiae contains transferases which catalyze the incorporation of N-acetylglucosamine from UDP-N-acetylglucosamine into a lipid fraction as well as into a protein fraction. The lipid fraction contains two alkali-stable lipids which can be separated on a silica G-60 column. The sugar moieties of these polyprenoid lipids are: N-acetylglucosamine and di-N-acetylchitobiose. The transfer of carbohydrate from isolated glycolipids to endogenous protein has been examined. After separation of protein and saccharide by hydrazinolysis and reacetylation only di-N-acetylchitobiose is found, and also when glycolipid containing only one N-acetylglucosamine is used as substrate. Maximum transfer of saccharides from glycolipids to protein is obtained at a Triton X-100 concentration of 1%. At this Triton X-100 concentration there is practically no transfer of N-acetylglucosamine from UDP-N-acetylglucosamine to the phosphorylated lipid. Therefore, when polyprenyl diphosphate N-acetyl[3H]-glucosamine is incubated together with UDP-N-acetyl[14C]glucosamine with the membrane fraction in the presence of 1% Triton X-100, a doubly labelled di-N-acetylchitobiose linked to lipid is formed with N-acetyl[14C]glucosamine at the non-reducing end of the chain.
D000117:Acetylglucosamine / Q000032:analysis / Q000378:metabolism*; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002855:Chromatography, Thin Layer; D005656:Fungal Proteins / Q000096:biosynthesis*; D005944:Glucosamine / Q000031:analogs & derivatives*; D006017:Glycolipids / Q000378:metabolism*; D007700:Kinetics; D008565:Membrane Proteins / Q000096:biosynthesis*; D011092:Polyethylene Glycols / Q000494:pharmacology; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000378:metabolism*; D014537:Uridine Diphosphate N-Acetylglucosamine / Q000378:metabolism
D005656:Fungal Proteins; D006017:Glycolipids; D008565:Membrane Proteins; D011092:Polyethylene Glycols; D014537:Uridine Diphosphate N-Acetylglucosamine; D005944:Glucosamine; D000117:Acetylglucosamine