High-speed single molecule imaging for evaluating the ERK transport mechanism locally around the nuclear pore complex
| Project/Area Number |
16K21625
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Multi-year Fund |
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| Research Field |
General anatomy (including histology/embryology)
Cell biology
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| Research Institution | Institute of Physical and Chemical Research |
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Principal Investigator |
Mouri Kazunari 国立研究開発法人理化学研究所, 生命システム研究センター, 研究員 (00567513)
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| Project Period (FY) |
2016-04-01 – 2018-03-31
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| Project Status |
Completed (Fiscal Year 2017)
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| Budget Amount *help |
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2017: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2016: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
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| Keywords | １分子計測 / ERK / 核膜孔通過 / FCS / ３次元１分子計測 / 蛍光相互相関分光法 / MAPK / 核膜孔 / 3次元1分子計測 / MAPKシグナル伝達 / 拡散係数計測 / 絶対濃度定量 / シグナル伝達 / 核膜孔輸送 / 1分子計測 |
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| Outline of Final Research Achievements |
Cells respond to external signals as a binary way. We have revealed that the phosphorylation level of ERK gradually increased depending on external signals, but the translocation of ERK behaved like a switch for them. The characteristics of this analog-to-digital conversion will be reflected in the enzyme reaction of ERK transport through nuclear pore complex (NPC). To verify this kinetics, we developed a new fluorescence correlation spectroscopy (FCS) using a conventional confocal laser scanning microscopy. We integrated this method and FRAP experiments, and enabled the estimation of enzymatic reaction rate constants of NPC in transporting ERK. Furthermore, in order to directly clarify the transport mechanism of ERK, we developed a single molecule imaging system using a total internal reflection fluorescence microscopy and realized a single molecule ERK imaging on NPC.
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Report
(3 results)
Research Products
(11 results)
