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Protein crystallization: from purified protein to diffraction-quality crystal

Nature Methods volume 5, pages 147–153 (2008)Cite this article

Abstract

Determining the structure of biological macromolecules by X-ray crystallography involves a series of steps: selection of the target molecule; cloning, expression, purification and crystallization; collection of diffraction data and determination of atomic positions. However, even when pure soluble protein is available, producing high-quality crystals remains a major bottleneck in structure determination. Here we present a guide for the non-expert to screen for appropriate crystallization conditions and optimize diffraction-quality crystal growth.

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Figure 1: The pipeline from gene to structure, illustrating the steps from target selection to structure determination and beyond.
Figure 2: Schematic illustration of a protein crystallization phase diagram.

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Acknowledgements

We acknowledge funding from the Engineering and Physical Sciences Research Council UK (EP/D501113/1) and the European Commission OptiCryst project LSHG-CT-2006-037793.

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Authors and Affiliations

  1. Naomi E. Chayen is in the Department of Biomolecular Medicine, Division of Surgery, Oncology, Reproductive Biology and Anaesthetics, Faculty of Medicine, Imperial College London, Sir Alexander Fleming Building, London SW7 2AZ, UK.,

    Naomi E Chayen

  2. Emmanuel Saridakis is at the Laboratory of Structural and Supramolecular Chemistry, Institute of Physical Chemistry, National Centre for Scientific Research “Demokritos”, Ag. Paraskevi, 15310 Athens, Greece.,

    Emmanuel Saridakis

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  1. Naomi E Chayen
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Correspondence to Naomi E Chayen.

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Supplementary Figures 1–2, Supplementary Note 1, Supplementary Protocols 1–2 (PDF 514 kb)

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Chayen, N., Saridakis, E. Protein crystallization: from purified protein to diffraction-quality crystal. Nat Methods 5, 147–153 (2008). https://doi.org/10.1038/nmeth.f.203

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