Abstract

An understanding of radiation damage effects suffered by biological samples during structural analysis using both X-rays and electrons is pivotal to obtain reliable molecular models of imaged molecules. This special issue on radiation damage contains six papers reporting analyses of damage from a range of biophysical imaging techniques. For X-ray diffraction, an in-depth study of multi-crystal small-wedge data collection single-wavelength anomalous dispersion phasing protocols is presented, concluding that an absorbed dose of 5 MGy per crystal was optimal to allow reliable phasing. For small-angle X-ray scattering, experiments are reported that evaluate the efficacy of three radical scavengers using a protein designed to give a clear signature of damage in the form of a large conformational change upon the breakage of a disulfide bond. The use of X-rays to induce OH radicals from the radiolysis of water for X-ray footprinting are covered in two papers. In the first, new developments and the data collection pipeline at the NSLS-II high-throughput dedicated synchrotron beamline are described, and, in the second, the X-ray induced changes in three different proteins under aerobic and low-oxygen conditions are investigated and correlated with the absorbed dose. Studies in XFEL science are represented by a report on simulations of ultrafast dynamics in protic ionic liquids, and, lastly, a broad coverage of possible methods for dose efficiency improvement in modalities using electrons is presented. These papers, as well as a brief synopsis of some other relevant literature published since the last Journal of Synchrotron Radiation Special Issue on Radiation Damage in 2019, are summarized below.

Highlights

  • There is already a substantial body of literature addressing various aspects of the radiation damage phenomena [see for example the 80 papers in special issues of the Journal of Synchrotron Radiation (JSR) arising from talks and posters given at the 2nd to 10th International Workshops on Radiation Damage to Biological Crystalline Samples, published in 2002, 2005, 2007, 2009, 2011, 2013, 2015, 2017 and 2019, respectively], which together provide a collected resource to researchers interested in the topic

  • Collecting X-ray crystallographic data from tiny macromolecular crystals is becoming more and more widespread thanks to increasingly intense synchrotron microfocus beamlines (Yamamoto et al, 2017), and is facilitated by dedicated programs, such as the automatic system for high-throughput structure analysis ZOO (Hirata et al, 2019). The latter supports small-wedge synchrotron crystallography (SWSX) that consists of collecting small-wedge sub-datasets from a multitude of microcrystals mounted in a cryo-loop that are merged into a full dataset

  • This optimum-dose value is in line with the point of diminishing returns for S-SAD phasing above which radiation-induced deterioration of data quality outweighs the gain in that data quality arising from increased multiplicity (Storm et al, 2017)

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Summary

Introduction

There is already a substantial body of literature addressing various aspects of the radiation damage phenomena [see for example the 80 papers in special issues of the Journal of Synchrotron Radiation (JSR) arising from talks and posters given at the 2nd to 10th International Workshops on Radiation Damage to Biological Crystalline Samples, published in 2002, 2005, 2007, 2009, 2011, 2013, 2015, 2017 and 2019, respectively], which together provide a collected resource to researchers interested in the topic. Factors affecting specific damage to particular amino acids (observed in electron density maps) have been investigated by Bhattacharyya and co-workers (Bhattacharyya et al, 2020) who sought to explain the differential radiation sensitivity of disulfide bonds observed in six different proteins.

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