| Budget Amount *help |
¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000)
Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2016: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
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| Outline of Final Research Achievements |
We developed new relaxation optimized SAIL amino acids for studying structural dynamics of large molecular proteins. Moreover, we also developed cost effective E. coli cellular expression systems, using auxotrophic mutants lacking certain enzymes in the biosynthetic pathways of various amino acids. From these results, we could successfully apply them to the 82 kDa MSG and about 1 MDa GroEL-GroES to observe the 13CH side chain moieties. By using high-pressure NMR experiments, we clearly detected structural fluctuations in core domain of MSG which could not be observed in conventional NMR experiments.
Unfortunately, we could not do the quantitative analysis for structural dynamics of MSG and GroEL-GroES due to the lower sensitivity of exchange signals. We will continue to improve and develop the SAIL-NMR methods to solve these problems.
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