| Budget Amount *help |
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2013: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
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| Outline of Final Research Achievements |
Tyrosine kinases are key protein for cellular signal transduction. Their abnormal activation is abundantly observed in various human cancers, and thought to be attractive therapeutic targets. NMR is a powerful tool for drug discovery. Its use is, however, quite limited due to availability of the isotope labeled tyrosine kinase by difficulty in production by bacterial expression system. In the present project, we developed the general method for production of the isotope labeled tyrosine kinase, and applied for evaluating conformational state of tyrosine kinase by NMR. Initially, we optimized promoter, fusion-tag protein, culturing condition and co-expression partner, and revealed that isotope labeled catalytic domain of eight tyrosine kinases can be successfully expressed by using E.coli expression system. Moreover, we identified that conformational state could be detected by using selective isotope labeled sample. The present result will contribute for NMR based drug discovery.
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