A simple adaptation to a protein crystallography station to facilitate difference X-ray scattering studies

Amit Sharma; Peter Berntsen; Rajiv Harimoorthy; Roberto Appio; Jennie Sjöhamn; Michael Järvå; Alexander Björling; Greger Hammarin; Sebastian Westenhoff; Gisela Brändén; Richard Neutze

doi:10.1107/S1600576719001900

A simple adaptation to a protein crystallography station to facilitate difference X-ray scattering studies

J Appl Crystallogr. 2019 Mar 28;52(Pt 2):378-386. doi: 10.1107/S1600576719001900. eCollection 2019 Apr 1.

Authors

Affiliations

¹ Department of Chemistry and Molecular Biology, University of Gothenburg, Box 462, 40530 Gothenburg, Sweden.
² Multidisciplinary Centre for Advanced Research and Studies, Jamia Millia Islamia, New Delhi 110025, India.
³ ARC Centre of Exellence in Advanced Molecular Imaging, Department of Chemistry and Physics, La Trobe Institute for Molecular Science, La Trobe University, Victoria 3086, Australia.
⁴ MAX IV Laboratory, Box 118, 221 00 Lund, Sweden.

Abstract

The X-ray crystallography station I911-2 at MAXLab II (Lund, Sweden) has been adapted to enable difference small- and wide-angle X-ray scattering (SAXS/WAXS) data to be recorded. Modifications to the beamline included a customized flow cell, a motorized flow cell holder, a helium cone, a beam stop, a sample stage and a sample delivery system. This setup incorporated external devices such as infrared lasers, LEDs and reaction mixers to induce conformational changes in macromolecules. This platform was evaluated through proof-of-principle experiments capturing light-induced conformational changes in phytochromes. A difference WAXS signature of conformational changes in a plant aqua-porin was also demonstrated using caged calcium.

Keywords: difference X-ray scattering; protein structural changes.

Grants and funding

This work was funded by Knut and Alice Wallenberg Foundation grants KAW 2012.0275 and KAW 2012.0284. Swedish Strategic Research Foundation grant SSF SRL 10-0036 to Richard Neutze. Swedish Research Council grants 2015-00560 and 349-2011-6485.