2013 Fiscal Year Final Research Report
Real-time observation of proton-pumping of membrane protein complexes using a novel high-sensitivity IR spectrometer
| Project/Area Number |
23685040
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| Research Category |
Grant-in-Aid for Young Scientists (A)
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| Allocation Type | Single-year Grants |
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| Research Field |
Chemistry related to living body
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| Research Institution | Institute of Physical and Chemical Research (2012-2013)
University of Hyogo (2011) |
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Principal Investigator |
KUBO Minoru 独立行政法人理化学研究所, 放射光科学総合研究センター, 研究員 (90392878)
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| Project Period (FY) |
2011-04-01 – 2014-03-31
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| Keywords | 赤外分光法 / 時間分解振動分光法 / ヘムタンパク質 / 膜タンパク質 / チトクロム酸化酵素 / プロトンポンプ / 一酸化窒素還元酵素 / フェムト秒レーザー |
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| Research Abstract |
This project aims to establish time-resolved IR spectroscopic techniques for observing reactions of membrane protein complexes under physiological conditions in real time. High-sensitivity IR spectrometer was constructed using a femtosecond IR laser as the light source. On the other hand, a flow-flash measurement system was developed to control reactions, which are triggered by the photolysis of caged compounds. Micro-channel flow cell was used to obtain a spectrum with minimal sample consumption. The flow-flash system was installed into time-resolved UV-visible absorption spectrometer and tested on NO reductase. We successfully observed the enzymatic reaction of this membrane protein with microsecond time resolution. Flow-flash time-resolved IR spectroscopy is ongoing.
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[Journal Article] Effective pumping-proton collection facilitated by a copper site (Cu_B) of bovine heart cytochrome c oxidase, revealed by a newly developed time-resolved infrared system2013
Author(s)
Kubo, M., Nakashima, S., Yamaguchi, S., Ogura, T., Mochizuki, M., Kang, J., Tateno, M., Shinzawa-Itoh, K., Kato, K., Yoshikawa, S
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Journal Title
J. Biol. Chem
Volume: Vol. 288 (Selected for JBC Bioenergetics Most-Read Articles)
Pages: 30259-30269
DOI
Peer Reviewed
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[Journal Article] Redox properties of a mononuclear copper(II)-superoxide complex2013
Author(s)
Tano, T., Okubo, Y., Kunishita, A., Kubo, M., Sugimoto, H., Fujieda, N., Ogura, T., Itoh, S
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Journal Title
Inorg. Chem
Volume: Vol. 52
Pages: 10431-10437
DOI
Peer Reviewed
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[Journal Article] Oxo-sulfido- and oxo-selenido-molybdenum(VI) complexes possessing dithiolene ligand related to the active sites of hydroxylases of molybdoenzymes : Low temperature preparation and characterization2013
Author(s)
Sugimoto, H., Tatemoto, S., Toyota, K., Kubo, M., Ogura, T., Itoh, S
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Journal Title
Chem. Commun
Volume: Vol. 49 (Selected for Cover Picture)
Pages: 4358-4360
DOI
Peer Reviewed
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[Journal Article] A new series of bis(ene-1,2-dithiolato)tungsten(IV), -(V), -(VI) complexes as reaction centre models of tungsten enzymes : preparation, crystal structures and spectroscopic properties2013
Author(s)
Sugimoto, H., Hatakeda, K., Toyota, K., Tatemoto, S., Kubo, M., Ogura, T., Itoh, S
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Journal Title
Dalton Trans
Volume: Vol. 42
Pages: 3059-3070
DOI
Peer Reviewed
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[Journal Article] Copper(I)-dioxygen reactivity in a sterically demanding tripodal tetradentate tren ligand. Formation and reactivity of a mononuclear copper(II) end-on superoxo complex2012
Author(s)
Kobayashi, Y., Okubo, K., Nomura, T., Kubo, M., Fujieda, N., Sugimoto, H., Fukuzumi, S., Goto, K., Ogura, T., Itoh, S
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Journal Title
Eur. J. Inorg. Chem
Volume: (29) (Selected for Cover Picture)
Pages: 4574-4578
DOI
Peer Reviewed
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[Journal Article] Chromium(IV)-peroxo complex formation and its nitric oxide dioxygenase reactivity2012
Author(s)
Yokoyama, A., Han, J. E., Cho, J., Kubo, M., Ogura, T., Siegler, M. A., Karlin, K. D., Nam, W
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Journal Title
J. Am. Chem. Soc
Volume: Vol. 134
Pages: 15269-15272
DOI
Peer Reviewed
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[Journal Article] Site-specific protein dynamics in communication pathway from sensor to signaling domain of oxygen sensor protein, HemAT-Bs : Time-resolved ultraviolet resonance Raman study2012
Author(s)
El-Mashtoly, S. F., Kubo, M., Gu, Y., Sawai, H., Nakashima, S., Ogura, T., Aono, S., Kitagawa, T
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Journal Title
J. Biol. Chem
Volume: Vol. 287
Pages: 19973-19984
DOI
Peer Reviewed
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[Journal Article] Chromium(V)-oxo and chromium(III)-superoxo complexes bearing a macrocyclic TMC ligand in hydrogen atom abstraction reactions2011
Author(s)
Cho, J., Woo, J., Han, J. E., Kubo, M., Ogura, T., Nam, W
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Journal Title
Chem. Sci
Volume: Vol. 2
Pages: 2057-2062
DOI
Peer Reviewed
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[Presentation] Development of a high-sensitivity time-resolved infrared spectrometer and its application to cytochrome c oxidase in aqueous solution2013
Author(s)
Kubo, M., Nakashima, S., Yamaguchi, S., Ogura, T., Mochizuki, M., Kang, J., Tateno, M., Shinzawa-Itoh, K., Kato, K., Yoshikawa, S
Organizer
International Conference on Structural Genomics-Structural Life Science (ICSG-2013-SLS)
Place of Presentation
Sapporo (Japan)
Year and Date
20130729-0801
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[Presentation] Real time observation of water-channel closure controlled by Cu_B in bovine cytochrome c oxidase using a novel high-sensitivity infrared system2013
Author(s)
Kubo, M., Nakashima, S., Yamaguchi, S., Ogura, T., Mochizuki, M., Kang, J., Tateno, M., Shinzawa-Itoh, K., Kato, K., Yoshikawa, S
Organizer
16th International Conference on Biological Inorganic Chemistry (ICBIC-16)
Place of Presentation
Grenoble (France)
Year and Date
20130722-26
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