Structure and mechanism of Helicobacter pylori fucosyltransferase. A basis for lipopolysaccharide variation and inhibitor design

J Biol Chem. 2007 Mar 30;282(13):9973-9982. doi: 10.1074/jbc.M610285200. Epub 2007 Jan 24.

Abstract

Helicobacter pylori alpha1,3-fucosyltransferase (FucT) is involved in catalysis to produce the Lewis x trisaccharide, the major component of the bacteria's lipopolysaccharides, which has been suggested to mimic the surface sugars in gastric epithelium to escape host immune surveillance. We report here three x-ray crystal structures of FucT, including the FucT.GDP-fucose and FucT.GDP complexes. The protein structure is typical of the glycosyltransferase-B family despite little sequence homology. We identified a number of catalytically important residues, including Glu-95, which serves as the general base, and Glu-249, which stabilizes the developing oxonium ion during catalysis. The residues Arg-195, Tyr-246, Glu-249, and Lys-250 serve to interact with the donor substrate, GDP-fucose. Variations in the protein and ligand conformations, as well as a possible FucT dimer, were also observed. We propose a catalytic mechanism and a model of polysaccharide binding not only to explain the observed variations in H. pylori lipopolysaccharides, but also to facilitate the development of potent inhibitors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Crystallography, X-Ray
  • Enzyme Inhibitors / chemical synthesis*
  • Fucosyltransferases / antagonists & inhibitors*
  • Fucosyltransferases / chemistry*
  • Fucosyltransferases / physiology
  • Helicobacter pylori / chemistry
  • Helicobacter pylori / enzymology*
  • Lipopolysaccharides / antagonists & inhibitors*
  • Lipopolysaccharides / biosynthesis
  • Lipopolysaccharides / chemistry*
  • Protein Conformation
  • Sequence Homology, Amino Acid

Substances

  • Enzyme Inhibitors
  • Lipopolysaccharides
  • Fucosyltransferases

Associated data

  • PDB/2NZW
  • PDB/2NZX
  • PDB/2NZY