| Project/Area Number |
15H03120
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Chemical biology
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| Research Institution | Tohoku University |
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Principal Investigator |
MIZUKAMI Shin 東北大学, 多元物質科学研究所, 教授 (30420433)
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| Co-Investigator(Renkei-kenkyūsha) |
KIKUCHI KAZUYA 大阪大学, 大学院工学研究科, 教授 (70292951)
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| Research Collaborator |
MATSUI YUSUKE 大阪大学, 大学院工学研究科, 大学院生
SATO RYOTA 大阪大学, 大学院工学研究科, 大学院生
AKAZAWA KAZUKI 大阪大学, 大学院工学研究科, 大学院生
SUZUKI SHUNSUKE 大阪大学, 大学院工学研究科, 大学院生
IMOTO TAKUMA 大阪大学, 大学院工学研究科, 大学院生
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| Project Period (FY) |
2015-04-01 – 2018-03-31
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| Project Status |
Completed (Fiscal Year 2017)
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| Budget Amount *help |
¥17,420,000 (Direct Cost: ¥13,400,000、Indirect Cost: ¥4,020,000)
Fiscal Year 2017: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2016: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2015: ¥6,630,000 (Direct Cost: ¥5,100,000、Indirect Cost: ¥1,530,000)
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| Keywords | 蛍光イメージング / タンパク質ラベル化 / Mg2+ / 蛋白質ラベル化 / 細胞内Mg2+ / 光制御 / 蛍光プローブ / 生細胞イメージング |
|---|
| Outline of Final Research Achievements |
We developed a fluorescent probe MGH with a chloroalkyl group, which can be labeled on a HaloTag. By labeling HaloTag expressed in living cells, MGH remained within living cells sufficiently even after 24 h of the treatment. Then, we attempted intracellular Mg2+ imaging during apoptosis, and visualized the free Mg2+ concentration increase after the apoptotic cell shrinkage. Various analyses indicated that the increase in intracellular free Mg2+ concentration during apoptosis was the result of release from Mg-ATP. We also developed fluorescent probes MGQ-1 and MGQ-2 that show high selectivity for Mg2+. The dissociation constants of MGQ-2 for Mg2+ and Ca2+were 0.27 mM and 1.5 mM, respectively, in buffer solution (pH 7.4), and MGQ-2 did not coordinate in the physiological concentration range of Mg-ATP.
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