| Project/Area Number |
16K08341
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Drug development chemistry
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| Research Institution | National Institute of Infectious Diseases |
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Principal Investigator |
Fukazawa Hidesuke 国立感染症研究所, 品質保証・管理部, 主任研究官 (10218878)
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| Project Period (FY) |
2016-04-01 – 2020-03-31
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| Project Status |
Completed (Fiscal Year 2019)
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| Budget Amount *help |
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2018: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2016: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
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| Keywords | チロシンキナーゼ / 細菌型チロシンキナーゼ / スクリーニング / タンパク質間相互作用 / BY-kinase |
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| Outline of Final Research Achievements |
Bacteria have tyrosine kinases that have no similarities to eukaryotic protein kinases. Although the functions of bacterial tyrosine kinases have not been fully revealed, their inhibitors are expected to become new antibacterial drugs because bacterial tyrosine kinases play essential roles in biosynthesis and transport of capsular polysaccharide and are closely related to pathogenicity. In this study, we successfully expressed active S. aureus tyrosine kinase in a human cell line. By fusing fluorescent protein tags to each of the catalytic subunit and regulatory subunit, we succeeded in visualizing the interaction between both subunits and revealed that the interaction and the kinase activation are strongly correlated. Various mutants were prepared, and sites important for enzyme activity and interaction were identified. We also established an inhibitor screening system for bacterial tyrosine kinases.
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| Academic Significance and Societal Importance of the Research Achievements |
細菌にもチロシンリン酸化酵素は存在し、病原性と深く関わっているが、真核生物のプロテインキナーゼとは類似性がなく、その阻害剤は選択性の高い新しい抗菌薬になることが期待されている。しかし簡便な測定法がなく、阻害剤開発は進んでいない。本研究では黄色ブドウ球菌のチロシンキナーゼをヒト培養細胞株に発現させ、酵素活性の検出と触媒－調節サブユニット間相互作用の可視化に成功した。この方法は従来法よりもはるかに簡便に酵素学的性質の解析や阻害剤探索を行うことが可能であり、新規抗菌物質の開発に役立つことが期待される。
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