| Budget Amount *help |
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2014: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2013: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
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| Outline of Final Research Achievements |
A simple technique for protein crystallization in ionically cross-linked polysaccharide (alginate and carrageenan) gel capsules has been developed for contactless handling of crystals in X-ray crystallography. This method is designed to reduce mechanical damage to crystals caused by physical contact between crystal and mount tool and by osmotic shock during various manipulations including cryoprotection, heavy-atom derivatization and ligand soaking experiments. In this study, ten test proteins could be successfully crystallized in gel capsules. Two complete diffraction data sets from crystals of proteins in gel capsules were collected at 100 K without removing the crystals, showing that the crystals had low mosaicities. To demonstrate heavy-atom derivatization, lysozyme crystals were successfully derivatized with Pt atoms within alginate gel capsules. These results suggest that gel capsules prevent serious damage to protein crystals during such experiments.
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