title: Inhibition of UDP-N-acetylglucosamine import into Golgi membranes by nucleoside monophosphates.
abstract: The specificity of the UDP-N-acetylglucosamine (UDP-GlcNAc) translocator for the binding of nucleoside monophosphates (NMPs) and nucleotide-sugars was examined in order to develop a quantitative understanding of how this enzyme recognizes its substrates and to provide a framework for development of novel drugs that target glycosylation. Competition studies reveal that tight binding requires a complete ribose ring and a 5'-phosphate. The enzyme is extremely tolerant to changes at the 3'-position, and the presence of 3'-F actually increases binding of the NMP to the enzyme. At the 2'-position, substitutions in the ribo configuration are well tolerated, although these same substitutions greatly diminish binding when present in the ara configuration. For the base, size appears to be the key feature for discrimination. The enzyme tolerates changing the C-4 oxygen of uridine to an amino group as well as substituting groups containing one or two carbons at C-5. However, substitution of groups containing three carbons at C-5, or exchange of the pyrimidine for a purine, greatly weakens binding to the translocator. Comparison of various UDP-sugars reveals that the UDP-GlcNAc translocator has lower affinity for UDP-N-acetylgalactosamine and UDP-glucose than for its cognate substrate and therefore indicates that this translocator requires both proper stereochemistry at C-4 and an aminoacetyl group at C-2. The impact of these observations on the design of more powerful nucleoside-based inhibitors of nucleotide-sugar import is discussed.
authors.affiliation: Department of Chemistry and Biochemistry, University of Colorado, Boulder 80309-0215, USA.
authors.forename: A J
authors.initials: AJ
authors.lastname: Traynor
authors.forename: E T
authors.initials: ET
authors.lastname: Hall
authors.forename: G
authors.initials: G
authors.lastname: Walker
authors.forename: W H
authors.initials: WH
authors.lastname: Miller
authors.forename: P
authors.initials: P
authors.lastname: Melançon
authors.forename: R D
authors.initials: RD
authors.lastname: Kuchta
mesh_terms: D000818:Animals; D001667:Binding, Competitive; D001692:Biological Transport / Q000187:drug effects; D002352:Carrier Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D006031:Glycosylation; D006056:Golgi Apparatus / Q000648:ultrastructure*; D007425:Intracellular Membranes / Q000378:metabolism*; D007700:Kinetics; D008099:Liver / Q000648:ultrastructure*; D009711:Nucleotides / Q000737:chemistry / Q000494:pharmacology*; D010710:Phosphates / Q000737:chemistry / Q000494:pharmacology; D011817:Rabbits; D012266:Ribose / Q000737:chemistry; D013329:Structure-Activity Relationship; D014537:Uridine Diphosphate N-Acetylglucosamine / Q000378:metabolism* / Q000494:pharmacology; D014539:Uridine Diphosphate Sugars / Q000378:metabolism / Q000494:pharmacology
chemical_list: D002352:Carrier Proteins; D009711:Nucleotides; D010710:Phosphates; D014539:Uridine Diphosphate Sugars; D014537:Uridine Diphosphate N-Acetylglucosamine; D012266:Ribose