| Budget Amount *help |
¥16,380,000 (Direct Cost: ¥12,600,000、Indirect Cost: ¥3,780,000)
Fiscal Year 2016: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000)
Fiscal Year 2015: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000)
Fiscal Year 2014: ¥6,110,000 (Direct Cost: ¥4,700,000、Indirect Cost: ¥1,410,000)
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| Outline of Final Research Achievements |
The so-called “Piston Movement Model” has been proposed in 2013, which assumes that the membrane domain of respiratory complex I (NADH-quinone oxidoreductase) undergoes large structural changes to pump protons across membrane. To verify this model by single-molecule analyses using functional molecular probes that can detect rotation or flick, we aimed to establish the methodology that enables the specific chemical modify of the enzyme. I succeeded in the specific modification (azidation) of Asp160 in 49 kDa subunit (Asp160-(CH2)3-N3), which is located in the quinone binding pocket of complex I, via ligand-directed tosyl（LDT）chemistry. I demonstrated that this azido group is able to serve as a “footing” for subsequent diverse chemical modifications via the so-called click chemistry.
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