Neutron Laue diffractometry with an imaging plate provides an effective data collection regime for neutron protein crystallography

Abstract

Neutron quasi-Laue diffraction data (2 A resolution) from tetragonal hen egg-white lysozyme were collected in ten days with neutron imaging plates. The data processing Laue software, LAUEGEN, developed for X-ray Laue diffractometry, was adapted for neutron diffractometry with a cylindrical detector. The data analysis software, X-PLOR, was modified and used for the refinement of hydrogen atoms, and the positions of 960 hydrogen atoms in the protein and 157 bound water molecules, were determined. Several examples are given of the methods used to identify hydrogen atoms and water molecules.

Enhanced visibility of hydrogen atoms by neutron crystallography on fully deuterated myoglobin

“…This situation is changing. Both the reliability of sources as well as advances in neutron detectors and data collection strategies (6) suggest that the technique will undergo a resurgence.…”

“…Even with large crystals, exposure times are long, normally in tens of minutes per degree of rotation, leading to data collection times of weeks or months. These difficulties are being addressed by improvements in instrumentation as well as in new methods of data collection (6). Two other technical problems are intrinsic to unlabeled samples.…”

A Neutron Laue Diffraction Study of Endothiapepsin: Implications for the Aspartic Proteinase Mechanism

“…The combination of a broad band-pass quasi-Laue geometry (e.g., λ ) 3.5 Å, dλ/λ ) 25%) with a novel cylindrical, neutron sensitive image plate detector that completely surrounds the sample provides 10-100-fold gains in efficiency compared with those of conventional monochromatic neutron diffractometers (21). This has made feasible studies of larger biological complexes and smaller crystals than was previously possible and has enabled the location of important hydrogen and/or deuterium atom positions in a number of systems at the resolutions typical of the majority of protein structure analyses (22)(23)(24)(25). This approach therefore offers great promise in the study and location of key hydrogen atom positions involved in enzymatic mechanisms, proton pumping, and shuttling processes and in protein-ligand bonding interactions.…”

Neutron crystallographic study on rubredoxin from Pyrococcus furiosus by BIX-3, a single-crystal diffractometer for biomacromolecules

“…In addition, we have developed a neutron-imaging plate (6)(7)(8)(9)(10)(11), which is a key component in our detector system, covering a large area with a high signal-to-noise ratio, high spatial accuracy, and good neutron-detection efficiency. Several studies in which our neutron-imaging plate detectors were used in neutron crystallography have been reported (12)(13)(14)(15)(16)(17)(18), including contributions from other groups who have adopted portions of our design (13,14,(16)(17)(18). All of these features have been incorporated in a neutron-imaging-plate-based diffractometer called BIX-3 (19)(20)(21) that has been built and installed at the JRR-3M reactor of the Japan Atomic Energy Research Institute.…”