Most high throughput structural proteomics centers use the sitting-drop method to obtain diffracting crystals for three-dimensional (3D) structure determination of biological macromolecules by x-ray crystallography. Although several robotic systems are available for dispensing the initial sitting-drop screening conditions, generally they are not used for optimization of crystallization conditions. This chapter describes a protocol for such automated systems, which permits easy construction of pH optimization grids using any desired fixed buffer set with varying ionic strengths directly dispensed into the crystallization plate.
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Acknowledgments
The authors acknowledge a grant from The Israel Ministry of Science, Culture and Sport to the ISPC, and the support of the Divadol Foundation, the Newman Foundation, and the European Commission Sixth Framework Research and Technological Development Programme SPINE2-COMPLEXES Project under contract No. 031220. The authors thank Israel Silman for his critical reading of the manuscript. J. L. Sussman is the Morton and Gladys Pickman Professor of Structural Biology.
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© 2008 Humana Press, a part of Springer Science+Business Media, LLC
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Meged, R., Dym, O., Sussman, J.L. (2008). High Throughput pH Optimization of Protein Crystallization. In: Kobe, B., Guss, M., Huber, T. (eds) Structural Proteomics. Methods in Molecular Biology™, vol 426. Humana Press. https://doi.org/10.1007/978-1-60327-058-8_27
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DOI: https://doi.org/10.1007/978-1-60327-058-8_27
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