Homogeneous batch micro-crystallization of proteins from ammonium sulfate
Open Access
- 26 January 2021
- journal article
- research article
- Published by International Union of Crystallography (IUCr) in Acta crystallographica. Section D, Structural biology
- Vol. 77 (2), 194-204
- https://doi.org/10.1107/s2059798320015454
Abstract
The emergence of X-ray free-electron lasers has led to the development of serial macromolecular crystallography techniques, making it possible to study smaller and more challenging crystal systems and to perform time-resolved studies on fast time scales. For most of these studies the desired crystal size is limited to a few micrometres, and the generation of large amounts of nanocrystals or microcrystals of defined size has become a bottleneck for the wider implementation of these techniques. Despite this, methods to reliably generate microcrystals and fine-tune their size have been poorly explored. Working with three different enzymes, L-aspartate α-decarboxylase, copper nitrite reductase and copper amine oxidase, the precipitating properties of ammonium sulfate were exploited to quickly transition from known vapour-diffusion conditions to reproducible, large-scale batch crystallization, circumventing the tedious determination of phase diagrams. Furthermore, the specific ammonium sulfate concentration was used to fine-tune the crystal size and size distribution. Ammonium sulfate is a common precipitant in protein crystallography, making these findings applicable to many crystallization systems to facilitate the production of large amounts of microcrystals for serial macromolecular crystallography experiments.Keywords
Funding Information
- H2020 Marie Skłodowska-Curie Actions (722687)
- Bundesministerium für Bildung, Wissenschaft und Forschung (05K16GU1)
- Federal Excellence Cluster Hamburg Centre for Ultrafast Imaging (EXC 1074)
- Biotechnology and Biological Sciences Research Council (BB/M022714/1)
This publication has 68 references indexed in Scilit:
- An Overview of Biological Macromolecule CrystallizationInternational Journal of Molecular Sciences, 2013
- Volume-conserving trans–cis isomerization pathways in photoactive yellow protein visualized by picosecond X-ray crystallographyNature Chemistry, 2013
- iMOSFLM: a new graphical interface for diffraction-image processing withMOSFLMActa crystallographica. Section D, Structural biology, 2011
- Femtosecond X-ray protein nanocrystallographyNature, 2011
- Exploring the Roles of the Metal Ions in Escherichia coli Copper Amine Oxidase,Biochemistry, 2010
- Patterns of protein–protein interactions in salt solutions and implications for protein crystallizationProtein Science, 2007
- Protein production by auto-induction in high-density shaking culturesProtein Expression and Purification, 2005
- Identification of Tyr58 as the proton donor in the aspartate-α-decarboxylase reactionChemical Communications, 2001
- REACTIVITY AND CRYOENZYMOLOGY OF ENZYMES IN THE CRYSTALLINE STATEAnnual Review of Biophysics and Bioengineering, 1977
- Zur Lehre von der Wirkung der SalzeNaunyn-Schmiedebergs Archiv für experimentelle Pathologie und Pharmakologie, 1888