European Biophysics Journal

, Volume 35, Issue 7, pp 601–609

A quasi-Laue neutron crystallographic study of d-xylose isomerase


    • Institut Laue Langevin
  • Edward H. Snell
    • Hauptman-Woodward Medical Research Institute
  • Mark J. van der Woerd
    • BAE Systems
  • Russell A. Judge
    • Abbott Laboratories
  • Dean A. A. Myles
    • Oak Ridge National Laboratory

DOI: 10.1007/s00249-006-0066-6

Cite this article as:
Meilleur, F., Snell, E.H., van der Woerd, M.J. et al. Eur Biophys J (2006) 35: 601. doi:10.1007/s00249-006-0066-6


The location of hydrogen atoms in enzyme structures can bring critical understanding of catalytic mechanism. However, whilst it is often difficult to determine the position of hydrogen atoms using X-ray crystallography even with subatomic (< 1.0 Å) resolution data available, neutron crystallography provides an experimental tool to directly localize hydrogen/deuterium atoms in biological macromolecules at resolution of 1.5–2.0 Å. d-Xylose isomerase (d-xylose ketol-isomerase, EC is a 43 kDa enzyme that catalyses the first reaction in the catabolism of d-xylose. Linearization and isomerization of d-xylose at the active site of d-xylose isomerase rely upon a complex hydrogen transfer. Neutron quasi-Laue data at 2.2 Å resolution were collected at room temperature on a partially deuterated Streptomyces rubiginosusd-xylose isomerase crystal using the LADI instrument at ILL with the objective to provide insight into the enzymatic mechanism. The neutron structure shows unambiguously that residue His 53 is doubly protonated at the active site of the enzyme. This suggests that the reaction proceeds through an acid catalyzed opening of the sugar ring, which is in accord with the mechanism suggested by Fenn et al. (Biochemistry 43(21): 6464–6474, 2004). This is the first report of direct observation of double protonation of His 53 and the first validation of the ring opening mechanism at the active site of d-xylose isomerase.

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© EBSA 2006