Stohrer, Claudia and Horrell, Sam and Meier, Susanne and Sans, Marta and von Stetten, David and Hough, Michael and Goldman, Adrian and Monteiro, Diana CF and Pearson, Arwen R (2021) Homogeneous batch micro-crystallization of proteins from ammonium sulfate. Acta Crystallographica Section D: Structural Biology, 77 (2). pp. 194-204. DOI https://doi.org/10.1107/s2059798320015454
Stohrer, Claudia and Horrell, Sam and Meier, Susanne and Sans, Marta and von Stetten, David and Hough, Michael and Goldman, Adrian and Monteiro, Diana CF and Pearson, Arwen R (2021) Homogeneous batch micro-crystallization of proteins from ammonium sulfate. Acta Crystallographica Section D: Structural Biology, 77 (2). pp. 194-204. DOI https://doi.org/10.1107/s2059798320015454
Stohrer, Claudia and Horrell, Sam and Meier, Susanne and Sans, Marta and von Stetten, David and Hough, Michael and Goldman, Adrian and Monteiro, Diana CF and Pearson, Arwen R (2021) Homogeneous batch micro-crystallization of proteins from ammonium sulfate. Acta Crystallographica Section D: Structural Biology, 77 (2). pp. 194-204. DOI 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.
Item Type: | Article |
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Uncontrolled Keywords: | Ammonium Sulfate; Crystallography, X-Ray; Macromolecular Substances; Proteins; microcrystals; batch crystallization; serial crystallography |
Divisions: | Faculty of Science and Health Faculty of Science and Health > Life Sciences, School of |
SWORD Depositor: | Unnamed user with email elements@essex.ac.uk |
Depositing User: | Unnamed user with email elements@essex.ac.uk |
Date Deposited: | 04 Oct 2024 10:42 |
Last Modified: | 30 Oct 2024 17:26 |
URI: | http://repository.essex.ac.uk/id/eprint/39328 |
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Filename: Homogeneous batch micro-crystallization of proteins from ammonium sulfate.pdf
Licence: Creative Commons: Attribution 4.0