Development of a dose-limiting data collection strategy for serial synchrotron rotation crystallography.

微小タンパク質結晶からの効率的な構造解析法 : 凍結した試料を回転させる「SS-ROX法」を確立、汎用化へ. 京都大学プレスリリース. 2017-01-12.Serial crystallography, in which single-shot diffraction images are collected, has great potential for protein microcrystallography. Although serial femtosecond crystallography (SFX) has been successfully demonstrated, limited beam time prevents its routine use. Inspired by SFX, serial synchrotron crystallography (SSX) has been investigated at synchrotron macromolecular crystallography beamlines. Unlike SFX, the longer exposure time of milliseconds to seconds commonly used in SSX causes radiation damage. However, in SSX, crystals can be rotated during the exposure, which can achieve efficient coverage of the reciprocal space. In this study, mercury single-wavelength anomalous diffraction (Hg-SAD) phasing of the luciferin regenerating enzyme (LRE) was performed using serial synchrotron rotation crystallography. The advantages of rotation and influence of dose on the data collected were evaluated. The results showed that sample rotation was effective for accurate data collection, and the optimum helical rotation step depended on multiple factors such as multiplicity and partiality of reflections, exposure time per rotation angle and the contribution from background scattering. For the LRE microcrystals, 0.25° was the best rotation step for the achievable resolution limit, whereas a rotation step larger than or equal to 1° was favorable for Hg-SAD phasing. Although an accumulated dose beyond 1.1 MGy caused specific damage at the Hg site, increases in resolution and anomalous signal were observed up to 3.4 MGy because of a higher signal-to-noise ratio

Constraining the Contribution of Galaxies and Active Galactic Nuclei to Cosmic Reionization

International audienceUnderstanding the detailed process of cosmic reionization is one of the remaining problems in astrophysics and cosmology. Here we construct a model of cosmic reionization that includes contributions from high-z galaxies and active galactic nuclei (AGNs) and calculate reionization and thermal histories with the model. To keep the model general and realistic, we vary the escape fraction of ionizing photons, f_esc, and the faint-end slope of the AGN luminosity function at high redshifts, αhz, within constraints from the observed cosmic star formation history and observed bright-end UV luminosity functions at z ≤ 6. Additionally, we model the spectral energy distribution (SED) of AGNs, which depends on the Eddington ratio and the black hole mass. By comparing the computed reionization histories with the observed H i fractions and the optical depth for Thomson scattering from Planck, we find that αhz > −1.5 and fesc < 0.15 are favoured when we employ the bright-end luminosity function obtained by Giallongo et al. Our result suggests that an AGN-dominated model with an abundance of faint AGNs as large as the estimate by Giallongo et al. is allowed only if the contribution from high-z galaxies is almost negligible, while a galaxy-dominant model is also allowed. We also find that the shape of the SED has a significant impact on the thermal history. Therefore it is expected that measurements of the thermal state of the intergalactic medium (IGM) will provide useful information on the properties of ionizing sources

Protein microcrystallography using synchrotron radiation

The progress in X-ray microbeam applications using synchrotron radiation is beneficial to structure determination from macromolecular microcrystals such as small in meso crystals. However, the high intensity of microbeams causes severe radiation damage, which worsens both the statistical quality of diffraction data and their resolution, and in the worst cases results in the failure of structure determination. Even in the event of successful structure determination, site-specific damage can lead to the misinterpretation of structural features. In order to overcome this issue, technological developments in sample handling and delivery, data-collection strategy and data processing have been made. For a few crystals with dimensions of the order of 10 µm, an elegant two-step scanning strategy works well. For smaller samples, the development of a novel method to analyze multiple isomorphous microcrystals was motivated by the success of serial femtosecond crystallography with X-ray free-electron lasers. This method overcame the radiation-dose limit in diffraction data collection by using a sufficient number of crystals. Here, important technologies and the future prospects for microcrystallography are discussed