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Figure 1
The setup and configuration of the counter-diffusion crystallization device, a JAXA crystallization box (JCB). (a) Parts of the crystallization device before assembly. (b) Outline of the assembled crystallization device for the microgravity experiment. It was mounted in an aluminium container, the Granada Crystallization Facility, and launched to the ISS. (c) Sample loading and crystallization-device setup. (i) Preparation of solutions. The protein solution (4.0 mg ml−1 human H-PGDS with or without 0.5 mM inhibitor in 150 mM sodium chloride, 15% PEG 6000, 5 mM dithiothreitol, 5 mM glutathione, 1% dioxane, 0.5 mM magnesium chloride and 20 mM Tris–HCl pH 8.0) and the precipitant solution (30% PEG 6000, 10 mM dithiothreitol, 10 mM glutathione, 1% dioxane and 1 mM magnesium chloride in 50 mM Tris–HCl pH 8.4) were prepared. The gel-tubes, which were polymerized agarose gels in a piece of plastic tubing, were incubated in 15% PEG 6000 solution containing 10 mM dithiothreitol, 10 mM glutathione, 2% dioxane, 1 mM magnesium chloride and 50 mM Tris–HCl pH 8.4 for 10 d before crystallization-device setup. (ii) Loading solutions and assembling the crystallization device. The protein solution was loaded into a capillary (1). The top of the capillary was tentatively sealed with clay and the gel-tube was plugged into the end of the capillary (2). The precipitant solution was loaded into the outer tube (3). The capillaries were inserted into the outer tube (4). The bottoms of the outer tubes were covered with caps and the top of the capillaries were completely sealed with epoxy adhesive (5).

ISSN: 2053-230X
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