Structure and function relationship of enzymes studied by single-molecule spectroscopy at liquid-helium temperature

2011 Fiscal Year Final Research Report

• Project Area: Molecular Science for Supra Functional Systems ? Development of Advanced Methods for Exploring Elementary Process
• Project/Area Number: 19056011
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Science and Engineering
• Research Institution: Tokyo Institute of Technology
• Principal Investigator: MATSUSHITA Michio 東京工業大学, 大学院・理工学研究科, 准教授 (80260032)
• Co-Investigator(Renkei-kenkyūsha): FUJIYOSHI Satoru 東京工業大学, 理工学研究科, 助教 (70371705)
• Project Period (FY): 2007 – 2011
• Keywords: 単一分子分光 / タンパク質 / 分光 / 低温 / 構造機能相関

Research Abstract

Physical and chemical properties of enzyme can be studied using a homogeneous solution of a purified sample of enzyme. However, in living cells, individual protein molecules of enzyme are in different locations to experience different environments, so that they function differently. For understanding enzymatic reactions on the basis of molecular science, observation of individual molecules is essential. In this study we realized spectroscopy of visible-fluorescent single protein molecules.

Research Products

• [Journal Article] Single-component reflecting objective for low-temperature imaging and spectroscopy of single nano objects (2011)
  • Author(s): M. Fujiwara, S. Fujiyoshi, M. Matsushita
  • Journal Title: Physics Procedia Volume: 13 Pages: 38
• [Journal Article] Single-protein study of photoresistance of pigment-protein complex in lipid bilayer (2011)
  • Author(s): D. Uchiyama, H. Hoshino, K. Otomo, T. Kato, K. Onda, A. Watanabe, H. Oikawa, S. Fujiyoshi, M. Matsushita, M. Nango, N. Watanabe, A. Sumino, T. Dewa
  • Journal Title: Chem. Phys. Lett. Volume: 511 Pages: 135
• [Journal Article] Reconstitution of bacterial photosynthetic unit in a lipid bilayer studied by single-molecule spectroscopy at 5 K (2011)
  • Author(s): D. Uchiyama, H. Oikawa, K. Otomo, M. Nango, T. Dewa, S. Fujiyoshi, M. Matsushita
  • Journal Title: Phys. Chem. Chem. Phys. Volume: 13 Pages: 11615
• [Journal Article] Structural Change of a Cofactor Binding Site of Flavoprotein Detected by Single-Protein Fluorescence Spectroscopy at 1.5 K (2011)
  • Author(s): S. Fujiyoshi, M. Hirano, M. Matsushita, M. Iseki, M. Watanabe
  • Journal Title: Phys. Rev. Lett. Volume: 106 Pages: 078101
• [Journal Article] タンパク質の構造変化と単一分子分光 (2011)
  • Author(s): 小井川浩之、藤芳暁、出羽毅久、南後守、松下道雄
  • Journal Title: 高分子 Volume: 60 (2) Pages: 80-81
• [Journal Article] Vibrational Microspectroscopy of Single Proteins (2010)
  • Author(s): S. Fujiyoshi, Y. Furuya, M. Iseki, M. Watanabe, M. Matsushita
  • Journal Title: J. Phys. Chem. Lett. Volume: 1 Pages: 2541-45
• [Journal Article] Single-component reflecting objective for ultraviolet imaging and spectroscopy at cryogenic temperature (2009)
  • Author(s): M. Fujiwara, S. Fujiyoshi, and M. Matsushita
  • Journal Title: J. Opt. Soc. Am. B Volume: 26 Pages: 1395-99
• [Journal Article] Visible fluorescence spectroscopy of single proteins at liquid-helium temperature (2008)
  • Author(s): S. Fujiyoshi, M. Fujiwara, and M. Matsushita
  • Journal Title: Phys. Rev. Lett. Volume: 100 Pages: 168101
• [Journal Article] How deep is the potential well confining a protein in a specific conformation? A single-molecule study on temperature dependence of conformational change between 5 and 18 K (2008)
  • Author(s): H. Oikawa, S. Fujiyoshi, T. Dewa, M. Nango, and M. Matsushita
  • Journal Title: J. Am. Chem. Soc. Volume: 130 Pages: 4580-81
• [Journal Article] 光合成アンテナ複合体の単一分子分光 (2008)
  • Author(s): 松下道雄
  • Journal Title: 生物物理 Volume: 48 (1) Pages: 35-40
• [Journal Article] Single-component reflecting objective for low-temperature spectroscopy in the entire visible region (2007)
  • Author(s): S. Fujiyoshi, M. Fujiwara, C. Kim, M. Matsushita, A. M. van Oijen, and J. Schmidt
  • Journal Title: App. Phys. Lett. Volume: 91 Pages: 051125
• [Presentation] 温度1.5Kの単一分子分光による可視蛍光タンパク質内のプロトン移動を伴った構造変化の観測 (2012)
  • Author(s): 藤原正規、平野充遥、渡辺正勝、伊関峰生、藤芳暁、松下道雄
  • Organizer: 日本化学会第92春季年会
  • Place of Presentation: 日吉
  • Year and Date: 20120325-28
• [Presentation] 中赤外光が誘起するタンパク質一分子の色素結合部位の構造変化 (2012)
  • Author(s): 大友康平、出羽毅久、松下道雄、藤芳暁
  • Organizer: 日本化学会第92春季年会
  • Place of Presentation: 日吉
  • Year and Date: 20120325-28
• [Presentation] 温度数ケルビンのタンパク質1分子分光装置の機械的安定性の評価 (2011)
  • Author(s): 日野原拓也、濱田裕紀、松下道雄、藤芳暁
  • Organizer: 第5回分子科学討論会
  • Place of Presentation: 札幌
  • Year and Date: 20110920-23
• [Presentation] 数Kにおける単一タンパク質分光を用いた可視蛍光タンパク質内のプロトン移動を伴った構造変化の観測 (2011)
  • Author(s): 藤原正規、平野充遥、渡辺正勝、伊関峰生、藤芳暁、松下道雄
  • Organizer: 第5回分子科学討論会
  • Place of Presentation: 札幌
  • Year and Date: 20110920-23
• [Presentation] Single-molecule spectroscopy of bacterial photosynthetic antenna complexes at liquid-helium temperature (2010)
  • Author(s): M. Matsushita
  • Organizer: The 70th Okazaki International Conference on "Molecular mechanism of photosynthetic energy conversion: the present research and future prospects"
  • Place of Presentation: Okazaki, Japan
  • Year and Date: 20101204-06
• [Presentation] 光合成アンテナ複合体の中赤外吸収スペクトルの一分子観測 (2010)
  • Author(s): 大友康平、出羽毅久、南後守、渡辺正勝、松下道雄、藤芳暁
  • Organizer: 第4回分子科学討論会
  • Place of Presentation: 大阪
  • Year and Date: 20100914-17
• [Presentation] 1.5Kにおける単一色素分子の蛍光スペクトルの時間変化の起源探究 (2010)
  • Author(s): 島内明理、櫻井敦教、平野充遥、藤芳暁、松下道雄
  • Organizer: 日本物理学会第65回年次大会
  • Place of Presentation: 岡山
  • Year and Date: 20100320-23
• [Presentation] Spectroscopy of single proteins at liquid-helium temperature (2010)
  • Author(s): M. Matsushita
  • Organizer: Symposium on "Physics and Chemistry of Coherently Controlled Quantum Systems"
  • Place of Presentation: Inuyama, Japan
  • Year and Date: 20100318-20
• [Presentation] 低温下における単一タンパク質分光のための反射対物レンズの開発 (2009)
  • Author(s): 藤原正規、藤芳暁、松下道雄
  • Organizer: 第3回分子科学討論会
  • Place of Presentation: 名古屋
  • Year and Date: 20090921-24
• [Presentation] 光活性化アデニル酸シクラーゼの単一タンパク質分光 (2009)
  • Author(s): 藤芳暁、平野充遥、松下道雄、伊関峰夫、渡辺正勝
  • Organizer: 第3回分子科学討論会
  • Place of Presentation: 名古屋
  • Year and Date: 20090921-24
• [Presentation] 低温の単一タンパク質分光の励起過程の検討 (2008)
  • Author(s): 藤芳暁、平野充遥、古屋陽、藤原正規、松下道雄、伊関峰夫、渡辺正勝
  • Organizer: 第2回分子科学討論会
  • Place of Presentation: 福岡
  • Year and Date: 20080924-27
• [Presentation] 顕微分光による光合成アンテナ複合体間のエネルギー移動の研究 (2008)
  • Author(s): 古屋内山大輔、藤芳暁、松下道雄、末守良春、出羽毅久、南後守
  • Organizer: 日本物理学会第63回年次大会
  • Place of Presentation: 東大阪
  • Year and Date: 20080322-26
• [Presentation] 1.5Kで使える反射対物レンズの球面収差及び色収差の改善 (2008)
  • Author(s): 藤原正規、藤芳暁、松下道雄
  • Organizer: 日本物理学会第63回年次大会
  • Place of Presentation: 東大阪
  • Year and Date: 20080322-26
• [Presentation] ニ光子蛍光による低温の単一タンパク質分光 (2008)
  • Author(s): 藤芳暁、松下道雄
  • Organizer: 日本物理学会第63回年次大会
  • Place of Presentation: 東大阪
  • Year and Date: 20080322-26
• [Presentation] Single-molecule spectroscopy of light-harvesting 2 complex at liquid-helium temperature (2007)
  • Author(s): H. Oikawa, S. Fujiyoshi, T. Dewa, M. Nango, and M. Matsushita
  • Organizer: The 2nd international workshop on "Photosynthetic Antennae and Coherent Phenomena"
  • Place of Presentation: Osaka, Japan
  • Year and Date: 20071214-16
• [Presentation] Structure of light-harvesting 2 complex from Rps. acidophila in lipid bilayer of DMPC: A single-molecule study at 5 K (2007)
  • Author(s): D. Uchiyama, H. Oikawa, S. Fujiyoshi, T. Dewa, M. Nango, and M. Matsushita
  • Organizer: The 2nd international workshop on "Photosynthetic Antennae and Coherent Phenomena"
  • Place of Presentation: Osaka, Japan
  • Year and Date: 20071214-16
• [Presentation] Spectroscopy of Single Proteins at Liquid Helium Temperature (2007)
  • Author(s): M. Matsushita and S. Fujiyoshi
  • Organizer: The 2nd International Workshop on "Materials Science and Nano-Engineering"
  • Place of Presentation: Awaji Island, Japan
  • Year and Date: 20071201-05
• [Presentation] Spectroscopy of Single Proteins at Liquid Helium Temperature (2007)
  • Author(s): M. Matsushita
  • Organizer: The 67th Okazaki International Conference on "Molecular science and chemical biology of biomolecular function"
  • Place of Presentation: Okazaki, Japan
  • Year and Date: 20071110-12
• [Presentation] 人工脂質膜中における光合成アンテナ複合体の会合体の分光 (2007)
  • Author(s): 内山大輔、古屋陽、藤芳暁、松下道雄、出羽毅久、南後守
  • Organizer: 日本物理学会第62回年次大会
  • Place of Presentation: 札幌
  • Year and Date: 20070921-24
• [Presentation] 可視蛍光をプローブとした低温の単一タンパク質分光 (2007)
  • Author(s): 藤芳暁、藤原正規、松下道雄
  • Organizer: 第1回分子科学討論会
  • Place of Presentation: 仙台
  • Year and Date: 20070917-20
• [Presentation] 二波長の励起光を同時に利用できる低温の単一タンパク質分光装置の開発 (2007)
  • Author(s): 平野充遥、藤原正規、藤芳暁、松下道雄、伊関峰生、渡辺正勝
  • Organizer: 第1回分子科学討論会
  • Place of Presentation: 仙台
  • Year and Date: 20070917-20
• [Remarks]
  • URL: http://www.molphys.phys.titech.ac.jp