Site-specific modification of respiratory complex I for single molecule analysis

2016 Fiscal Year Final Research Report

• Project/Area Number: 26292060
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Bioorganic chemistry
• Research Institution: Kyoto University
• Principal Investigator: Miyoshi Hideto 京都大学, (連合)農学研究科(研究院), 教授 (20190829)
• 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)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Keywords: 生物制御化学 / 呼吸鎖複合体-I / 化学修飾 / ユビキノン

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.

Free Research Field

生物有機化学