title: Synthesis of aryl azide derivatives of UDP-GlcNAc and UDP-GalNAc and their use for the affinity labeling of glycosyltransferases and the UDP-HexNAc pyrophosphorylase.
abstract: The chemical synthesis and utilization of two photoaffinity analogs, 125I-labeled 5-[3-(p-azidosalicylamido)-1-propenyl]-UDP-GlcNAc and -UDP-GalNAc, is described. Starting with either UDP-GlcNAc or UDP-GalNAc, the synthesis involved the preparation of the 5-mercuri-UDP-HexNAc and then attachment of an allylamine to the 5 position to give 5-(3-amino)allyl-UDP-HexNAc. This was followed by acylation with N-hydroxysuccinimide p-aminosalicylic acid to form the final product, i.e., 5-[3-(p-azidosalicylamido)-1-propenyl]-UDP-GlcNAc or UDP-GalNAc. These products could then be iodinated with chloramine T to give the 125I-derivatives. Both the UDP-GlcNAc and the UDP-GalNAc derivatives reacted in a concentration-dependent manner with a highly purified UDP-HexNAc pyrophosphorylase, and both specifically labeled the subunit(s) of this protein. The labeling of the protein by the UDP-GlcNAc derivative was inhibited in dose-dependent fashion by either unlabeled UDP-GlcNAc or unlabeled UDP-GalNAc. Likewise, labeling with the UDP-GalNAc probe was blocked by either UDP-GlcNAc or UDP-GalNAc. The UDP-GlcNAc probe also specifically labeled a partially purified preparation of GlcNAc transferase I.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences, Little Rock, Arkansas, 72205, USA.
authors.forename: Y
authors.initials: Y
authors.lastname: Zeng
authors.forename: Y
authors.initials: Y
authors.lastname: Shabalin
authors.forename: T
authors.initials: T
authors.lastname: Szumilo
authors.forename: I
authors.initials: I
authors.lastname: Pastuszak
authors.forename: R R
authors.initials: RR
authors.lastname: Drake
authors.forename: A D
authors.initials: AD
authors.lastname: Elbein
mesh_terms: D000345:Affinity Labels / Q000138:chemical synthesis*; D000818:Animals; D001386:Azides; D016695:Glycosyltransferases / Q000032:analysis*; D009713:Nucleotidyltransferases / Q000032:analysis*; D010777:Photochemistry; D012457:Salicylamides / Q000138:chemical synthesis*; D012459:Salicylates; D013552:Swine; D014531:Uridine Diphosphate Galactose / Q000031:analogs & derivatives* / Q000138:chemical synthesis; D014532:Uridine Diphosphate Glucose / Q000031:analogs & derivatives* / Q000138:chemical synthesis; D014536:Uridine Diphosphate N-Acetylgalactosamine / Q000031:analogs & derivatives*; D014537:Uridine Diphosphate N-Acetylglucosamine / Q000031:analogs & derivatives*
chemical_list: C101284:5-(3-(4-azido-3,5-diiodosalicylamido)-1-propenyl)-uridine diphosphate N-acetylglucosamine; C101285:5-(3-(4-azido-3,5-diiodosalicylamido-)-1-propenyl)-uridine diphosphate N-acetylgalactosamine; D000345:Affinity Labels; D001386:Azides; D012457:Salicylamides; D012459:Salicylates; D014531:Uridine Diphosphate Galactose; D014537:Uridine Diphosphate N-Acetylglucosamine; C063873:4-azidosalicylic acid; D014536:Uridine Diphosphate N-Acetylgalactosamine; D016695:Glycosyltransferases; D009713:Nucleotidyltransferases; C023226:UDPacetylglucosamine pyrophosphorylase; D014532:Uridine Diphosphate Glucose