期刊
PHILOSOPHICAL TRANSACTIONS OF THE ROYAL SOCIETY B-BIOLOGICAL SCIENCES
卷 369, 期 1647, 页码 -出版社
ROYAL SOC
DOI: 10.1098/rstb.2013.0590
关键词
X-ray crystallography; X-ray emission spectroscopy; X-ray free-electron laser; metalloenzymes; water oxidation; photosystem II
类别
资金
- NIH [GM055302, P41GM103393, GM095887, GM102520]
- Office of Science, Office of Basic Energy Sciences (OBES), Division of Chemical Sciences, Geosciences and Biosciences (CSGB) of the Department of Energy (DOE) [DE-AC02-05CH11231]
- LBNL Laboratory Directed Research and Development award (DOE) [DE-AC02-05CH11231]
- Human Frontier Research grant [RGP0063/2013]
- Alexander von Humboldt Foundation
- Ruth L. Kirschstein National Research Service Award [F32GM100595]
- DOE Office of Basic Energy Sciences, Chemical Sciences Division [DE-AC02-76SF00515]
- Human Frontiers Science Project Award [RPG005/2011]
X-ray free-electron lasers (XFELs) open up new possibilities for X-ray crystallographic and spectroscopic studies of radiation-sensitive biological samples under close to physiological conditions. To facilitate these new X-ray sources, tailored experimental methods and data-processing protocols have to be developed. The highly radiation-sensitive photosystem II (PSII) protein complex is a prime target for XFEL experiments aiming to study the mechanism of light-induced water oxidation taking place at a Mn cluster in this complex. We developed a set of tools for the study of PSII at XFELs, including a new liquid jet based on electrofocusing, an energy dispersive von Hamos X-ray emission spectrometer for the hard X-ray range and a high-throughput soft X-ray spectrometer based on a reflection zone plate. While our immediate focus is on PSII, the methods we describe here are applicable to a wide range of metalloenzymes. These experimental developments were complemented by a new software suite, cctbx.xfel. This software suite allows for near-real-time monitoring of the experimental parameters and detector signals and the detailed analysis of the diffraction and spectroscopy data collected by us at the Linac Coherent Light Source, taking into account the specific characteristics of data measured at an XFEL.
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