TIM crystals grown by capillary counterdiffusion: Statistical evidence of quality improvement in microgravity

Evrard, Christine;Maes, Dorninique;Zegers, Ingrid;Declercq, Jean-Paul;Van De Weerdt, Ceile;et.al.
(2007) 11th International Conference on the Crystallization of Biological Macromolecules — Location: Quebec City(Canada) (16.August.2006)

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Authors
  • Evrard, Christine
    Author
  • Maes, Dorninique
    Author
  • Zegers, Ingrid
    Author
  • Declercq, Jean-PaulUCLouvain
    Author
  • Vanhee, CelineUCLouvain
    Author
  • Van De Weerdt, Ceile
    Author
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Abstract
The capillary counterdiffusion method is a very efficient crystallization technique for obtaining high-quality protein crystals. This technique requires a convection-free environment, which can be achieved using either gelled solutions, very thin capillaries, or microgravity conditions. To study the influence of a convection-free environment on protein crystal quality and to evaluate two different experimental implementations to achieve it, we have made a comparative analysis of crystals grown by capillary counterdiffusion in agarose, a convective-free environment on Earth, and crystals grown in microgravity at the International Space Station. Thermotoga maritima triose phosphate isomerase (TIM) was chosen as a model for this study. The statistical analysis reveals a significant improvement for the crystals grown in microgravity in terms of their R-merge, B-value, and mosaicity, but the statistical evidence is insufficient to show a similar benefit for the resolution and mean intensity parameters. These results are quite surprising because it is known that, unlike gels, the noisy microgravity scenario offered by the ISS cannot sustain a convection-free environment on the time scale of days required for protein crystallization experiments.
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Citations

Evrard, C., Maes, D., Zegers, I., Declercq, J.-P., Vanhee, C., Martial, J., Wyns, L., & Van De Weerdt, C. (2007). TIM crystals grown by capillary counterdiffusion: Statistical evidence of quality improvement in microgravity. Crystal Growth & Design, 7(11), 2161-2166. https://doi.org/10.1021/cg700687t (Original work published 2007)