Site-specific modification of respiratory complex I for single molecule analysis
| Project/Area Number |
26292060
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Partial Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Bioorganic chemistry
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| Research Institution | Kyoto University |
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Principal Investigator |
Miyoshi Hideto 京都大学, (連合)農学研究科(研究院), 教授 (20190829)
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| Co-Investigator(Renkei-kenkyūsha) |
MURAI Masatoshi 京都大学, 大学院農学研究科, 助教 (80543925)
FURUIKE Shou 大阪医科大学, 医学部, 講師 (60392875)
FUJIWARA Takahiro 京都大学, 物質細胞統合システム拠点, 講師 (80423060)
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| Project Period (FY) |
2014-04-01 – 2017-03-31
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| Project Status |
Completed (Fiscal Year 2016)
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| 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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| Keywords | 生物制御化学 / 呼吸鎖複合体-I / 化学修飾 / ユビキノン / ミトコンドリア呼吸鎖 / トシル化学 / 複合体-I / ミトコンドリア / 呼吸鎖 / 呼吸鎖酵素 |
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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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Report
(4 results)
Research Products
(12 results)
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[Journal Article] Specific methylation of Asp160 (49 kDa subunit) located inside the quinone binding cavity of bovine mitochondrial complex I2016
Author(s)
Murai, M., Inaoka, H., Masuya, T., Aburaya, S., Aoki, W., and Miyoshi, H.
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Journal Title
Biochemistry
Volume: 55
Issue: 23
Pages: 3189-3197
DOI
Related Report
Peer Reviewed / Acknowledgement Compliant
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