| Project/Area Number |
15K12753
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Chemical biology
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| Research Institution | University of Tsukuba |
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Principal Investigator |
KITA Masaki 筑波大学, 数理物質系, 准教授 (30335012)
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| Project Period (FY) |
2015-04-01 – 2017-03-31
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| Project Status |
Completed (Fiscal Year 2016)
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| Budget Amount *help |
¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2015: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
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| Keywords | 天然物ケミカルバイオロジー / 標的分子同定 / 質量分析法 / 光親和性プローブ / LA-LDI MS |
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| Outline of Final Research Achievements |
To establish new binding mode analysis methods of target biomacromolecules, we aimed to develop photoaffinity probes that can covalently bind to target molecules and efficiently analyze their fragment peptides by using label-assisted laser desorption/ionization mass spectrometry (LA-LDI MS). We found that 6-amidopyrene (apy) derivatives were highly detectable by the LDI MS instrument. An apy-conjugated photoaffinity derivative of an antitumor natural product, aplyronine A, was then synthesized. By the irradiation of UV light (365 nm), this probe efficiently photoreacted with solvent molecules and form covalent bonds. Furthermore, actin was quantitatively labeled with an apy NHS ester probe of aplyronine A, and the sequence of an apy-labeled peptide was established by MS/MS analysis.
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