Crystal structure of A. aeolicus LpxC with bound product suggests alternate deacetylation mechanism

Proteins. 2015 Sep;83(9):1706-19. doi: 10.1002/prot.24856. Epub 2015 Aug 1.

Abstract

UDP-3-O-acyl-N-acetylglucosamine deacetylase (LpxC) is the first committed step to form lipid A, an essential component of the outer membrane of Gram-negative bacteria. As it is essential for the survival of many pathogens, LpxC is an attractive target for antibacterial therapeutics. Herein, we report the product-bound co-crystal structure of LpxC from the acheal Aquifex aeolicus solved to 1.6 Å resolution. We identified interactions by hydroxyl and hydroxymethyl substituents of the product glucosamine ring that may enable new insights to exploit waters in the active site for structure-based design of LpxC inhibitors with novel scaffolds. By using this product structure, we have performed quantum mechanical modeling on the substrate in the active site. Based on our results and published experimental data, we propose a new mechanism that may lead to a better understanding of LpxC catalysis and inhibition.

Keywords: LpxC; QM/MM; deacetylase; dynamics; lipid A; lipopolysaccharide; mechanism; natural product; transition state; zinc.

MeSH terms

  • Acetylation
  • Amidohydrolases / chemistry*
  • Amidohydrolases / metabolism
  • Bacteria / enzymology*
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / metabolism
  • Biocatalysis
  • Catalytic Domain
  • Crystallography, X-Ray
  • Enzyme Inhibitors / chemistry
  • Enzyme Inhibitors / metabolism
  • Glucosamine / chemistry
  • Glucosamine / metabolism
  • Lipid A / biosynthesis
  • Models, Molecular
  • Molecular Structure
  • Protein Binding
  • Protein Structure, Tertiary*
  • Quantum Theory
  • Substrate Specificity
  • Zinc / chemistry
  • Zinc / metabolism

Substances

  • Bacterial Proteins
  • Enzyme Inhibitors
  • Lipid A
  • Amidohydrolases
  • UDP-3-O-acyl-N-acetylglucosamine deacetylase
  • Zinc
  • Glucosamine