European Biophysics Journal

, Volume 35, Issue 7, pp 577–583

Comparison of hydrogen determination with X-ray and neutron crystallography in a human aldose reductase–inhibitor complex

Authors

  • M. P. Blakeley
    • EMBL
  • A. Mitschler
    • IGBMC, CNRS/INSERM/ULP
  • I. Hazemann
    • IGBMC, CNRS/INSERM/ULP
  • F. Meilleur
    • ILL
  • D. A. A. Myles
    • Oak Ridge National Laboratory
    • IGBMC, CNRS/INSERM/ULP
Article

DOI: 10.1007/s00249-006-0064-8

Cite this article as:
Blakeley, M.P., Mitschler, A., Hazemann, I. et al. Eur Biophys J (2006) 35: 577. doi:10.1007/s00249-006-0064-8

Abstract

Protonation states determination by neutron (2.2 Å at room temperature) and X-ray (0.66 Å at 100 K) crystallographic studies were compared for a medium size enzyme, human aldose reductase (MW = 36 kDa), complexed with its NADP+ coenzyme and a selected inhibitor of therapeutic interest. The neutron resolution could be achieved only with the ab initio fully deuterated protein and the subsequent crystallization in D2O of the complex. We used the largest good-quality crystal (1.00 × 0.67 × 0.23 mm, i.e. volume of 0.15 mm3) that we were able to grow so far. Both studies enable the determination of protonation states, with a clear advantage for neutrons in the case of less-ordered atoms (B > 5 Å2). Hydrogen atoms are best determined by a complementary analysis of the Fourier maps obtained from both methods.

Copyright information

© EBSA 2006