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Complementarity of neutron, XFEL and synchrotron crystallography for defining the structures of metalloenzymes at room temperature

DOI: 10.1107/S2052252522006418 DOI Help

Authors: Tadeo Moreno-Chicano (University of Essex) , Leiah M. Carey (North Carolina State University) , Danny Axford (Diamond Light Source) , John H. Beale (Diamond Light Source) , R. Bruce Doak (Max Planck Institute for Medical Research) , Helen M. E. Duyvesteyn (University of Oxford) , Ali Ebrahim (University of Essex; Diamond Light Source) , Robert W. Henning (University of Chicago; Argonne National Laboratory) , Diana C. F. Monteiro (Hauptman-Woodward Medical Research Institute) , Dean A. Myles (Oak Ridge National Laboratory) , Shigeki Owada (Japan Synchrotron Radiation Research Institute) , Darren A. Sherrell (Diamond Light Source) , Megan L. Straw (University of Essex) , Vukica Šrajer (University of Chicago; Argonne National Laboratory) , Hiroshi Sugimoto (RIKEN SPring-8 Center) , Kensuke Tono (Japan Synchrotron Radiation Research Institute) , Takehiko Tosha (RIKEN SPring-8 Center) , Ivo Tews (University of Southampton) , Martin Trebbin (Hauptman-Woodward Medical Research Institute; State University of New York at Buffalo) , Richard W. Strange (University of Essex) , Kevin L. Weiss (Oak Ridge National Laboratory) , Jonathan A. R. Worrall (University of Essex) , Flora Meilleur (North Carolina State University) , Robin L. Owen (Diamond Light Source) , Reza A. Ghiladi (North Carolina State University) , Michael A. Hough (University of Essex; Diamond Light Source)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Iucrj , VOL 9

State: Published (Approved)
Published: September 2022
Diamond Proposal Number(s): 14493

Open Access Open Access

Abstract: Room-temperature macromolecular crystallography allows protein structures to be determined under close-to-physiological conditions, permits dynamic freedom in protein motions and enables time-resolved studies. In the case of metalloenzymes that are highly sensitive to radiation damage, such room-temperature experiments can present challenges, including increased rates of X-ray reduction of metal centres and site-specific radiation-damage artefacts, as well as in devising appropriate sample-delivery and data-collection methods. It can also be problematic to compare structures measured using different crystal sizes and light sources. In this study, structures of a multifunctional globin, dehaloperoxidase B (DHP-B), obtained using several methods of room-temperature crystallographic structure determination are described and compared. Here, data were measured from large single crystals and multiple microcrystals using neutrons, X-ray free-electron laser pulses, monochromatic synchrotron radiation and polychromatic (Laue) radiation light sources. These approaches span a range of 18 orders of magnitude in measurement time per diffraction pattern and four orders of magnitude in crystal volume. The first room-temperature neutron structures of DHP-B are also presented, allowing the explicit identification of the hydrogen positions. The neutron data proved to be complementary to the serial femtosecond crystallography data, with both methods providing structures free of the effects of X-ray radiation damage when compared with standard cryo-crystallography. Comparison of these room-temperature methods demonstrated the large differences in sample requirements, data-collection time and the potential for radiation damage between them. With regard to the structure and function of DHP-B, despite the results being partly limited by differences in the underlying structures, new information was gained on the protonation states of active-site residues which may guide future studies of DHP-B.

Journal Keywords: serial femtosecond crystallography; serial synchrotron crystallography; neutron crystallography; XFELs; room temperature; metalloenzymes

Diamond Keywords: Enzymes

Subject Areas: Biology and Bio-materials

Diamond Offline Facilities: XFEL-Hub
Instruments: I24-Microfocus Macromolecular Crystallography

Added On: 27/07/2022 14:21


Discipline Tags:

Structural biology Life Sciences & Biotech

Technical Tags:

Diffraction Serial Synchrotron Crystallography (SSX)