X-ray structure of perdeuterated diisopropyl fluorophosphatase (DFPase): perdeuteration of proteins for neutron diffraction
The signal‐to‐noise ratio is one of the limiting factors in neutron macromolecular crystallography. Protein perdeuteration, which replaces all H atoms with deuterium, is a method of improving the signal‐to‐noise ratio of neutron crystallography experiments by reducing the incoherent scattering of th...
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Published in | Acta crystallographica. Section F, Structural biology and crystallization communications Vol. 66; no. 4; pp. 379 - 385 |
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Main Authors | , , , , , , , |
Format | Journal Article |
Language | English |
Published |
5 Abbey Square, Chester, Cheshire CH1 2HU, England
International Union of Crystallography
01.04.2010
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Subjects | |
Online Access | Get full text |
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Summary: | The signal‐to‐noise ratio is one of the limiting factors in neutron macromolecular crystallography. Protein perdeuteration, which replaces all H atoms with deuterium, is a method of improving the signal‐to‐noise ratio of neutron crystallography experiments by reducing the incoherent scattering of the hydrogen isotope. Detailed analyses of perdeuterated and hydrogenated structures are necessary in order to evaluate the utility of perdeuterated crystals for neutron diffraction studies. The room‐temperature X‐ray structure of perdeuterated diisopropyl fluorophosphatase (DFPase) is reported at 2.1 Å resolution. Comparison with an independently refined hydrogenated room‐temperature structure of DFPase revealed no major systematic differences, although the crystals of perdeuterated DFPase did not diffract neutrons. The lack of diffraction is examined with respect to data‐collection and crystallographic parameters. The diffraction characteristics of successful neutron structure determinations are presented as a guideline for future neutron diffraction studies of macromolecules. X‐ray diffraction to beyond 2.0 Å resolution appears to be a strong predictor of successful neutron structures. |
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Bibliography: | istex:99E051F1525D2737E5857692EE4AC5A40757DD7C ArticleID:AYF2TB5020 ark:/67375/WNG-PLGFSC1K-8 ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 ObjectType-Article-2 ObjectType-Feature-1 |
ISSN: | 1744-3091 1744-3091 |
DOI: | 10.1107/S1744309110004318 |