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A mechanism-inspired UDP-N-acetylglucosamine pyrophosphorylase inhibitor

Research output: Contribution to journalJournal articlepeer-review

E-pub ahead of print
  • Olwale G. Raimi
  • Ramon Hurtado-Guerrero
  • Vladimir Borodkin
  • Andrew Ferenbach
  • Mick Urbaniak
  • Michael A J Ferguson
  • Daan M. F. van Aalten
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<mark>Journal publication date</mark>24/03/2020
<mark>Journal</mark>RSC Chemical Biology
Issue number1
Volume2020
Number of pages13
Pages (from-to)13-25
Publication StatusE-pub ahead of print
Early online date24/03/20
<mark>Original language</mark>English

Abstract

UDP-N-acetylglucosamine pyrophosphorylase (UAP1) catalyses the last step in eukaryotic biosynthesis of uridine diphosphate-N-acetylglucosamine (UDP-GlcNAc), converting UTP and GlcNAc-1P to the sugar nucleotide. Gene disruption studies have shown that this gene is essential in eukaryotes and a possible antifungal target, yet no inhibitors of fungal UAP1 have so far been reported. Here we describe the crystal structures of substrate/product complexes of UAP1 from Aspergillus fumigatus that together provide snapshots of catalysis. A
structure with UDP-GlcNAc, pyrophosphate and Mg2+ provides the first Michaelis complex trapped for this class of enzyme, revealing the structural basis of the previously reported Mg2+ dependence and direct observation of
pyrophosphorolysis. We also show that a highly conserved lysine mimics the role of a second metal observed in structures of bacterial orthologues. A mechanism-inspired UTP α,β-methylenebisphosphonate analogue (meUTP) was designed and synthesized and was shown to be a micromolar inhibitor of the enzyme. The mechanistic insights and inhibitor described here will facilitate future studies towards the discovery of small molecule inhibitors of this currently unexploited potential antifungal drug target.