International Journal of Microgravity Science and Application
Online ISSN : 2188-9783
Essential Techniques for Growing High-quality Protein Crystals for Structural Analysis in Microgravity
Koji INAKASachiko TAKAHASHIHiroaki TANAKA
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JOURNAL OPEN ACCESS

2017 Volume 34 Issue 1 Pages 340104-

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Abstract
For successful protein crystallization in microgravity environment, the following four points are essential: high-quality protein samples and well-optimized crystallization conditions with high reproducibility, crystallization conditions which are optimized for counter-diffusion method, and crystallization conditions which enhance microgravity effects. To meet these points, we have a strategy that we first evaluate the protein samples and crystallization conditions by charge-density calculation, SDS-PAGE, Native-PAGE, high-performance ion-exchange column chromatography, DLS and preliminary crystallization experiment by vapor-diffusion method. Then according to the results of these evaluation, we make a trial of improvement of the protein samples and crystallization conditions. If the results of the evaluation are consistent with the charge density, good protein crystals grow in most cases. For the crystals grown in JAXA PCG (protein crystal growth) space experiment, harvesting and cryocooling of the crystals from capillaries require some skills. In this review, we introduce these techniques for users who have already participated or will participate in JAXA PCG.
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© 2017 The Japan Society of Microgravity Application
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