Synthesis of UDP-N-[1-14C]acetyl D-glucosamine and UDP-N-[1-14C]acetyl-D-galactosamine from [1-14C]acetate.
Procedures for the preparation of UDP-N-[1-14C]acetyl-D-glucosamine and UDP-N-[1-14C]acetyl-D-galactosamine with very high specific activities are described. The overall yield based on the amount of [1-14C]acetate used is greater than 80%. The N-acetyl-D-glucosamine-alpha-1-phosphate used in this synthesis is prepared by phosphorylation of tetraacetyl-D-N-acetylglucosamine with crystalline phosphoric acid. N-acetyl-D-glucosamine-alpha-1-phosphate is then deacetylated in anhydrous hydrazine with hydrazine sulfate as a catalyst. D-glucosamine-alpha-1-phosphate is N-acetylated with [14C]acetate using N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline as the coupling agent. The acetylated product is coverted to the UDP derivative with yeast UDP-N-acetyl-D-glucosamine pyrophosphorylase. UDP-N-[1-14C]acetylgalactosamine is prepared by acetylation of UDP-galactosamine using [1-14C]acetate and N-ethoxy-carbonyl-2-ethoxy-1,2-dihydroquinoline. UDP-galactosamine is prepared enzymatically using galactokinase and galactose-1-phosphate uridyltransferase. The labeled products, isolated and characterized by ion-exchange and paper chromatography, were active as substrates in glycosyl transferase systems.
Department of Biochemistry, University of Georgia, Athens 30602, USA.
A K
AK
Rao
J
J
Mendicino
D019342:Acetic Acid; D000818:Animals; D002250:Carbon Radioisotopes; D002623:Chemistry Techniques, Analytical / Q000379:methods; D016695:Glycosyltransferases / Q000378:metabolism; D066298:In Vitro Techniques; D007202:Indicators and Reagents; D011804:Quinolines; D014536:Uridine Diphosphate N-Acetylgalactosamine / Q000138:chemical synthesis*; D014537:Uridine Diphosphate N-Acetylglucosamine / Q000138:chemical synthesis*
D002250:Carbon Radioisotopes; D007202:Indicators and Reagents; D011804:Quinolines; D014537:Uridine Diphosphate N-Acetylglucosamine; C000470:EEDQ; D014536:Uridine Diphosphate N-Acetylgalactosamine; D016695:Glycosyltransferases; D019342:Acetic Acid