生体関連分子の機能発現に対する溶媒効果の分子論的研究

• Project/Area Number: 15076211
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Science and Engineering
• Research Institution: Fukuoka University
• Principal Investigator: 山口 敏男 Fukuoka University, 理学部, 教授 (70158111)
• Co-Investigator(Kenkyū-buntansha): 吉田 亨次 福岡大学, 理学部, 助教 (00309890) ; 李 相男 福岡大学, 理学部, 研究員 (40248472)
• Project Period (FY): 2003 – 2007
• Project Status: Completed (Fiscal Year 2007)
• Budget Amount: ¥58,700,000 (Direct Cost: ¥58,700,000); Fiscal Year 2007: ¥5,900,000 (Direct Cost: ¥5,900,000); Fiscal Year 2006: ¥5,900,000 (Direct Cost: ¥5,900,000); Fiscal Year 2005: ¥12,800,000 (Direct Cost: ¥12,800,000); Fiscal Year 2004: ¥16,300,000 (Direct Cost: ¥16,300,000); Fiscal Year 2003: ¥17,800,000 (Direct Cost: ¥17,800,000)
• Keywords: タンパク質のフォールディング / 溶媒効果 / 疎水性-親水性バランス / アミロイド線維 / X線・中性子回折 / 溶媒クラスター / フルオロアルコール / β-ラクトグロブリン / アミロイド形成 / タンパク質 / X線・中性子散乱 / シミュレーション / アルコール-水混合溶液 / ジメチルスルホキシド-水混合溶液

Research Abstract

1.アルコール添加により形成されるβ-ラクトグロブリン(BLG)ゲルの構造とダイナミクスを中性子小角散乱および中性子スピンエコーによって研究した。エタノール濃度が20体積%まではBLGはモノマーで存在しているが、30体積%を超えると会合した。これは、BLGがα-ヘリックス-リッチ構造に変化するエタノール組成と一致した。また、TFEについても同様に、会合状態と二次構造変化の間でよい相関が得られた。エタノール、TFEのいずれの揚合もアルコール添加とともに、BLGの運動は急激に遅くなった。会合体のダイナミクスについても二次構造変化とよい相関があることを示した。
2・希薄BLGの水、エタノールー水、ならびにTFE-水各混合溶媒の非干渉性中性子散スペクトルを測定した。スペクトルは局所的な速いモードとび依存性を示す遅いモードに分離された。純水中より混合溶媒中では、二つのモードの緩和は遅く、平均自乗変位は小さいことが示された。の結果は混合溶媒中では、純水中に比べてBLGの取り得るコンフォメーションが制限されることを示しており、BLGの最安定構造に到達できず、その結果、ミスフォールドを生じるというアルコール変性についての一つの機構を提案した。
3.我々がデザインした人工タンパク質Amyloidgenesin-Met(以下AG-Met)について、蛍光タンパクGFPを融合させたAGMet-GFPを細胞内に導入し発現させたところ.蛍光顕微鏡観察で胞質中にドット状に会合したタンパク質がみられた.このものは、チオフラビンTで染色さることから、アミロイド様線維であることが分かった。しかし、顕著な細胞毒性が見られかったことから、このアミロイド繊維は,直接細胞死に関係のないことが示唆された。

Research Products

• [Journal Article] Structure and dynamic properties of liquids confined in MCM-41mesopores (2007)
  • Author(s): Yamaguchi, T., Yoshida, K., Smimov, P., Takamuku, T., Kittaka, S.,Takahara, T., Kuroda, Y. & Bellissent-Funel. M.-C.
  • Journal Title: Eur.Phys.J.Special Topics 141 Pages: 19-27
• [Journal Article] Liquid structure of acetic acid-water and trifluoroacetic acid-water miχturesIstudied by large-angle X-ray scattering andNMR (2007)
  • Author(s): Takamuku, T., Kyoshoin, Y・, Noguchi, H. Kusano, S., Yamaguchi, T.
  • Journal Title: J.Phys.Chem.B 111 Pages: 9270-9280
• [Journal Article] Interlayer water molecules in vanadium pentoxide hydrate.LX.Anisotropictranclational diffusion leading to anisotropic ac conductivity (2007)
  • Author(s): Kittaka, S., Yamaguchi, T.Bellisssent-Funel M.-C.
  • Journal Title: J.Chem.Phys. 127
• [Journal Article] モデル小ペプチドのアルコール誘起α-ヘリックス形成における溶媒環境の役割 (2007)
  • Author(s): 藤永丈晴、中村智、Krishtal Sergey, 吉田亨次、李相男、金澤健治、根本直、山口敏男、
  • Journal Title: 福岡大学理学集報 37・1 Pages: 23-31
• [Journal Article] Hydration water in dynamics of a hydrated β-lactoblubulin (2007)
  • Author(s): Yoshida, K., Yamaguchi, T., Bellissent-Funel, M.-C. Longeville, S.
  • Journal Title: Eur.Phys.J.Special Topics 141 Pages: 223-226
• [Journal Article] Hydrogen bonding in methanol studied by infrared spectroscopy (2007)
  • Author(s): Wu, X, ., Chen, Y., Yamaguchi, T.
  • Journal Title: J.Mol.Spectr. 246 Pages: 187-191
• [Journal Article] Hydration water in dynamics of a hydrated β-lactoblubulin (2007)
  • Author(s): Yoshida, K., Yamaguchi, T., Bellissent-Funel, M.-C.Longeville, S.
  • Journal Title: Eur. J. Phys. Special Topics 141 Pages: 19-27
• [Journal Article] Enthalpy and interfacial free energy changes of water capillary condensed in mesoporous silica, MCM-41 and SBA-15 (2006)
  • Author(s): Kittaka, S., Ishimaru, S., Kuranishi, M., Matsuda T., Yamaguchi, T
  • Journal Title: Physical Chemistry Chemical Physics 8 Pages: 3223-3231
• [Journal Article] Thermal properties and mixing state of ethylene glycol-water binary solutions by calorimetry, large-angle X-ray scattering, and small-angle neutron scattering (2006)
  • Author(s): Matsugami, M., Takamuku, T., Otomo T., Yamaguchi, T.
  • Journal Title: The Journal of Physical Chemistry A 110(25) Pages: 12372-12379
• [Journal Article] Amyloid formation and cytotoxicity of a de novo designed protein, amyloidgenesin, and its analogs (2006)
  • Author(s): Okada, T., Lee, S., Yoshida, K., Yamaguchi, T., Nakashima, M., Takahashi, M.
  • Journal Title: Peptide Science Pages: 217-220
  • NAID: 10017675682
• [Journal Article] ^<17>O NMR relaxation study of dynamics of water molecules in aqueous mixtures of methanol, ethanol, and 1-propanol over a temperature range of 283-403 K (2006)
  • Author(s): K, Yoshida, K., Kitajo, A., Yamaguchi, T.
  • Journal Title: Journal of Molecular Liquids 125 Pages: 158-163
• [Journal Article] Mode of Interaction of amphiphilic α-helical peptide with phosphatidylcholines at the air-water interface (2006)
  • Author(s): Nakahara, H., Nakamura, S., Hiranita, T., Kawasaki, H., Lee, S., Sugihara, G., Shibata.O.
  • Journal Title: Langmuir 22(3) Pages: 1182-1192
• [Journal Article] X-ray diffraction studies on methanol-water, ethanol-water, and 2-propanol-water mixtures at low temperatures. (2005)
  • Author(s): T.Takamuku, T., Saisho, K., Nozawa, S., Yamaguchi, T.
  • Journal Title: J.Mol.Liq. 119(1-3) Pages: 133-146
• [Journal Article] Neutron diffraction study on microinhomogeneities in ethanol-water mixtures (2005)
  • Author(s): Yamaguchi, T., Takamuku, T., Soper, A.K
  • Journal Title: J.Neutron Res. 13(1-3) Pages: 129-133
• [Journal Article] Structure and dynamics of halogenoethanol-water mixtures studied by large-angle x-ray scattering, small-angle neutron scattering, and NMR relaxation (2005)
  • Author(s): Takamuku, T., Kumai, T., Yoshida, K., Otomo, T., Yamaguchi, T.
  • Journal Title: J.Phys.Chem.A 109(34) Pages: 7667-7676
• [Journal Article] Neutron scattering study on dynamics of water molecules in MCM-41.2. Determination of translational diffusion coefficient. (2005)
  • Author(s): Takahara, S., Sumiyama, N., Kittaka, S., Yamaguchi, T., Bellissent-Funel, M.-C.
  • Journal Title: J.Phys.Chem.B 109(22) Pages: 11231-11239
• [Journal Article] Replica-exchange molecular dynamics simulation of small peptide in water and in ethanol. (2005)
  • Author(s): Yoshida, K., Yamaguchi, T., Okamoto, Y.
  • Journal Title: Chem.Phys.Lett 412 Pages: 280-284
• [Journal Article] 水と生体分子が織り成す生命現象:たんぱく質と中性子散乱 (2005)
  • Author(s): 山口敏男, 片岡幹雄
  • Journal Title: 現代化学 6月号 Pages: 55-59
• [Journal Article] Structure of 1-propanol-water mixtures investigated by large-angle x-ray scattering technique (2004)
  • Author(s): Takamuku.T., Maruyama, H., Watanabe, K., Yamaguchi, T
  • Journal Title: J.Solution Chem. 33 Pages: 641-660
• [Journal Article] Liquid structure of 1-propanol by molecular dynamics simulation and x-ray scattering (2004)
  • Author(s): Akiyama, I., Ogawa, M., Takase, K., Takamuku, T., Yamaguchi, T., Ohtori, N.
  • Journal Title: J.Solution Chem. 33 Pages: 797-809
• [Journal Article] Structure of aqueous gallium (III) bromide solutions over a temperature range 80-333 K by Raman spectroscopy, x-absorption fine structure, and x-ray diffraction (2004)
  • Author(s): Smirnov, P., Wakita, H., Nomura, M., Yamaguchi, T.
  • Journal Title: J.Solution Chem. 33 Pages: 903-922
• [Journal Article] Cluster structure in helix-promoting hexafluoro- iso- propanol - water mixtures (2004)
  • Author(s): Yoshida, K., Yamaguchi, T., Bowron, D., Finney, J.
  • Journal Title: J.Neutron Res. 12 Pages: 305-309
• [Journal Article] Organotin (IV) complexes of polyhydroxyalkyl carboxylic acids and some related ligands (2004)
  • Author(s): Szorcsik, A., Lagy, L., Gyurcsik, B., Vanko, G., Kramer, R., Vertes, A., Yamaguchi, T., Yoshida, K.
  • Journal Title: J.Radioanal.Nucl.Chem.. 260 Pages: 459-469
• [Journal Article] Acoustic phonon dynamics in liquid CCl_4 (2004)
  • Author(s): Kamiyama, T., Hosokawa, S., Baron, A.Q.R., Tsutsui, S., Yoshida, K., Pilgrim, W.C., Kiyanagi, Y., Yamaguchi, T.
  • Journal Title: J.Phys.Soc.Jpn. 73 Pages: 1615-1618
  • NAID: 110001954861
• [Presentation] On the Role of Solvent Environment in Alcohol-induced a-Helix Formation of Model Peptides (2007)
  • Author(s): T.Yamaguchi, T.Fujinaga, S.Nakamura, S.Krishtal, K.Yoshida, S.Lee, K.Kanazawa, T.Nemoto,
  • Organizer: 30th International Conference on Solution CHemistry
  • Place of Presentation: Perth,Australia
  • Year and Date: 2007-07-19
• [Patent(Industrial Property Rights)] Artificial pulmonary surfactants
  • Inventor(s): Lee, S., Yukitake K, Nakamura, Y
• [Publications] Yoshida, K., Yamaguchi, T., Adachi, T., Otomo, T., Matsuo, D., Takamuku, T., Nishi, N.: "Structure and dynamics of hexafluoroisopropanol-water mixtures by x-ray diffraction, small-angle neutron scattering, NMR spectroscopy, and mass spectroscopy"J.Chem.Phys.. 119. 6132-6142 (2003)
• [Publications] Takamuku, T., Matsuo, D., Tabata, M., Yamaguchi, T., Nishi, N.: "Structure of aqueous mixtures of N,N- dimethylacetamide studied by infrared spectroscopy, X-ray diffraction, and mass spectroscopy."J.Phys.Chem.. 107. 6070-6078 (2003)
• [Publications] Takamuku, T., Nakamizo, A., Tabata, M., Yoshida, K., Yamaguchi, T., Otomo, T.: "Large-angle x-ray scattering, small-angle neutron scattering, and NMR relaxation studies on mixing states of 1,4-dioxane-water, 1,3-dioxane-water, and tetrahydrofuran-water mixtures"J.Mol.Liq.. 103-104. 143-159 (2003)
• [Publications] 丸山浩和, 高椋利幸, 山口敏男, 橘高茂治, 高原周一: "X線回折法によるMCM-41細孔中に閉じ込められたメタノールの構造解析(2003)X線分析の進歩"X線分析の進歩. 34. 227-239 (2003)
• [Publications] Ellerby, H.M, Lee, S., Ellerby, L.M., Chen, S., Kiyota, T., del Rio, G., Sugihara, G., Sun, Y., Bredesen, D.E., Arap, W., Pasqualini, R.: "An artificially designed pore-forming protein with anti-tumor effects"J.Biol.Chem.. 278. 35311-35316 (2003)
• [Publications] Furuya, T., Kiyota, T., Lee, S., Inoue, T., Sugihara, G., Logvinova, A., Goldsmith, P., Ellerby, H.M.: "Nanotubules formed by highly hydrophobic amphiphilic alpha-helical peptides and natural phospholipids"Biophys.J.. 84. 1950-1959 (2003)