Crystallization Optimum Solubility Screening: using crystallization results to identify the optimal buffer for protein crystal formation

Acta Crystallographica. Section F, Structural Biology and Crystallization Communications
Bernard CollinsRebecca Page

Abstract

An optimal solubility screen is described that uses the results of crystallization trials to identify buffers that improve protein solubility and, in turn, crystallization success. This screen is useful not only for standard crystallization experiments, but also can easily be implemented into any high-throughput structure-determination pipeline. As a proof of principle, the predicted novel-fold protein AF2059 from Archaeoglobus fulgidus, which was known to precipitate in most buffers and particularly during concentration experiments, was selected. Using the crystallization results of 192 independent crystallization trials, it was possible to identify a buffer containing 100 mM CHES pH 9.25 that significantly improves its solubility. After transferring AF2059 into this ;optimum-solubility' buffer, the protein was rescreened for crystal formation against these same 192 conditions. Instead of extensive precipitation, as observed initially, it was found that 24 separate conditions produced crystals and the exchange of AF2059 into CHES buffer significantly improved crystallization success. Fine-screen optimization of these conditions led to the production of a crystal suitable for high-resolution (2.2 A) structure determination.

References

Aug 24, 2002·Proceedings of the National Academy of Sciences of the United States of America·Scott A LesleyRaymond C Stevens
Apr 30, 2003·Journal of Structural Biology·W William Wilson
Jun 5, 2003·Acta Crystallographica. Section D, Biological Crystallography·Rebecca PageScott A Lesley
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Citations

Jun 15, 2007·Extremophiles : Life Under Extreme Conditions·Francis E Jenney, Michael W W Adams
Jun 15, 2007·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Jhe-Le TuShan-Ho Chou
Mar 24, 2012·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Janet NewmanFrank von Delft
Oct 13, 2015·Acta Crystallographica. Section F, Structural Biology Communications·Marko RisticJanet Newman
Mar 29, 2016·Molecular Pharmaceutics·Ashlesha S Raut, Devendra S Kalonia
Jul 9, 2013·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Chen Yan ZhangPeng Shang

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