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

Protonation states determination by neutron (2.2 A at room temperature) and X-ray (0.66 A 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 res...

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Bibliographic Details
Published in:European biophysics journal 2006-09, Vol.35 (7), p.577-583
Main Authors: Blakeley, M P, Mitschler, A, Hazemann, I, Meilleur, F, Myles, D A A, Podjarny, A
Format: Article
Language:English
Subjects:
Aldehyde Reductase
Aldehyde Reductase - antagonists & inhibitors
Aldehyde Reductase - chemistry
Aldehyde Reductase - metabolism
Binding Sites
Biochemistry, Molecular Biology
Crystallization
Crystallography, X-Ray
Deuterium
Deuterium - chemistry
Humans
Hydrogen
Hydrogen - chemistry
Hydrogen Bonding
Life Sciences
Models, Molecular
Molecular biology
Molecular Structure
NADP
NADP - metabolism
Neutron Diffraction
Neutrons
Protein Conformation
Protons
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Summary:Protonation states determination by neutron (2.2 A at room temperature) and X-ray (0.66 A 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 (1.00x0.67x0.23 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 A2). Hydrogen atoms are best determined by a complementary analysis of the Fourier maps obtained from both methods.
ISSN:0175-7571
1432-1017
DOI:10.1007/s00249-006-0064-8