Two-dimensional crystallization conditions of human leukotriene C4 synthase 
requiring adjustment of a particularly large combination of specific parameters

Zhao, G.; Johnson, M. C.; Schnell, J. R.; Kanaoka, Y.; Haase, W.; Irikura, D.; Lam, B. K.; Schmidt-Krey, I.

doi:10.1016/j.jsb.2009.11.002

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Two-dimensional crystallization conditions of human leukotriene C₄ synthase requiring adjustment of a particularly large combination of specific parameters

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Haase, W.
Department of Structural Biology, Max Planck Institute of Biophysics, Max Planck Society;
Department of Physiology, Max Planck Institute of Biophysics, Max Planck Society;

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Citation

Zhao, G., Johnson, M. C., Schnell, J. R., Kanaoka, Y., Haase, W., Irikura, D., et al. (2010). Two-dimensional crystallization conditions of human leukotriene C₄ synthase requiring adjustment of a particularly large combination of specific parameters. Journal of Structural Biology, 169(3), 450-454. doi:10.1016/j.jsb.2009.11.002.

Cite as: https://hdl.handle.net/11858/00-001M-0000-0024-D6D0-A

Abstract

Human leukotriene C(4) synthase (LTC(4)S) forms highly ordered two-dimensional (2D) crystals under specific reconstitution conditions. It was found that control of a larger number of parameters than is usually observed for 2D crystallization of membrane proteins was necessary to induce crystal formation of LTC(4)S. Here, we describe the parameters that were optimized to yield large and well-ordered 2D crystals of LTC(4)S. Careful fractioning of eluates during the protein purification was essential for obtaining crystals. While the lipid-to-protein ratio was critical in obtaining order, four parameters were decisive in inducing growth of crystals that were up to several microns in size. To obtain a favorable diameter, salt, temperature, glycerol, and initial detergent concentration had to be controlled with great care. Interestingly, several crystal forms could be grown, namely the plane group symmetries of p2, p3, p312, and two different unit cell sizes of plane group symmetry p321.