X-ray and UV radiation-damage-induced phasing using synchrotron serial crystallography.

Nicolas Foos,Henry N. Chapman,Olof Svensson,Robin Schubert,Carolin Seuring,Max H. Nanao,Anja Burkhardt,Alke Meents

doi:10.1107/s2059798318001535

Abstract

Specific radiation damage can be used to determine phases de novo from macromolecular crystals. This method is known as radiation-damage-induced phasing (RIP). One limitation of the method is that the dose of individual data sets must be minimized, which in turn leads to data sets with low multiplicity. A solution to this problem is to use data from multiple crystals. However, the resulting signal can be degraded by a lack of isomorphism between crystals. Here, it is shown that serial synchrotron crystallography in combination with selective merging of data sets can be used to determine high-quality phases for insulin and thaumatin, and that the increased multiplicity can greatly enhance the success rate of the experiment.

Highlights

Radiation induces many changes in macromolecular crystals
Each data set acquired from both thaumatin and insulin microcrystals in the MeshAndCollect workflow was merged using CODGAS to obtain complete data sets
The increasing number of sub-data sets used to obtain a full data set might be due to degradation of the individual sub-data-set quality owing to nonspecific radiation damage, i.e. more sub-data sets are required for equivalent data quality

Summary

Introduction

Radiation induces many changes in macromolecular crystals. Amongst these is a reduction in occupancy or the movement of atoms, which is referred to as specific radiation damage. Specific radiation damage can be of major concern to practitioners of macromolecular crystallography (MX), but in some cases such damage can be used to determine phases experimentally (Ravelli et al, 2003, 2005; Zwart et al, 2004; Banumathi et al, 2004; Weiss et al, 2004; Schiltz et al, 2004; Ramagopal et al, 2005; de Sanctis & Nanao, 2012; de Sanctis et al, 2016) This technique is called radiation-damage-induced phasing (RIP) and, by analogy to single isomorphous replacement (SIR), two data sets are used to calculate differences in structure factors (between damaged and less damaged states).

Methods

Results

Conclusion