X-ray structure of perdeuterated diisopropyl fluorophosphatase (DFPase): Perdeuteration of proteins for neutron diffraction

Marc Michael Blum, Stephen J. Tomanicek, Harald John, B. Leif Hanson, Heinz Rüterjans, Benno P. Schoenborn, Paul Langan, Julian C.H. Chen

Research output: Contribution to journalArticlepeer-review

15 Scopus citations

Abstract

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 crystallo-graphic 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.

Original languageEnglish
Pages (from-to)379-385
Number of pages7
JournalActa Crystallographica Section F: Structural Biology and Crystallization Communications
Volume66
Issue number4
DOIs
StatePublished - 2010
Externally publishedYes

Keywords

  • Diisopropyl fluorophosphatase
  • Perdeuteration

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