Protein crystallization by capillary counterdiffusion for applied crystallographic structure determination |
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Authors: | Ng Joseph D Gavira José A García-Ruíz Juan M |
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Institution: | Department of Biological Sciences and the Laboratory for Structural Biology, University of Alabama at Huntsville, Huntsville, AL 35899, USA. ngj@email.uah.edu |
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Abstract: | Counterdiffusion crystallization in capillary is a very simple, cost-effective, and practical procedure for obtaining protein crystals suitable for X-ray data analysis. Its principles have been derived using well-known concepts coupling the ideas of precipitation and diffusion mass transport in a restricted geometry. The counterdiffusion process has been used to simultaneously screen for optimal conditions for protein crystal growth, incorporate strong anomalous scattering atoms, and mix in cryogenic solutions in a single capillary tube. The crystals obtained in the capillary have been used in situ for X-ray analysis. The implementation of this technique linked to the advancement of current crystallography software leads to a powerful structure determination method consolidating crystal growth, X-ray data collection, and ab initio phase determination into one without crystal manipulation. We review the historical progress of counterdiffusion crystallization, its application to X-ray crystallography, and ongoing tool development for high-throughput protein structure determination. |
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