1998 Fiscal Year Annual Research Report

タンパク質の動的構造研究のためのX線溶液散乱解析システムの開発

Research Project

Project/Area Number	09558093
Research Institution	Yokohama City University
Principal Investigator	佐藤衛横浜市立大学, 大学院・総合理学研究科, 教授 (60170784)
Co-Investigator(Kenkyū-buntansha)	雨宮慶幸東京大学, 工学部・総合試験所, 助教授 (70151131) 片岡幹雄奈良先端科学技術大学院大学, 物質創成科学研究科, 教授 (30150254)
Keywords	X線溶液散乱 / X線小角散乱 / 時分割測定 / 動的構造解析 / タンパク質の溶液構造
Research Abstract	佐藤は、昨年度に開発したストップトフロー混合装置を改良した試料槽を用いて、牛肝臓のプロテアソームおよびその活性化因子のX線溶液散乱測定を行ない、両者の複合体のマクロ構造を明らかにするとともに、両者が1:1の化学量論比で複合体を形成して機能することを明らかにした。両者の複合体形成については、すでに電子顕微鏡で1:2の化学量論比で形成されることが示されているが、1:2モデルでは説明できないタンパク質分解の機構が1:1モデルでは説明できることなど、プロテアソームの構造と機能の相関研究に新たな事実を提供した。片岡は、様々な変性状態などの非天然構造や合成ポリペプチドの取る構造をX線溶液散乱法で特徴づけ、非天然構造の有力な研究手段であることを示した。さらに、バクテリオロドプシンの光反応中間体のより詳細な構造解析のために、重原子置換法を応用し、7Å分解能の回折データを用いて投影図上に重原子の位置変化をマップできることを示し、重原子置換法を用いたX線溶液散乱法が二次元結晶配列をした膜蛋白質の構造研究に広く応用できることを示した。また、雨宮は、蛍光体と縮小型光ファイバーをCCDに組み合わせたモジュールをモザイク状に並べたアレイ状CCD型X線検出器を開発し、一号機として2×2個のアレイ状CCD型X線検出器(有効面積:144×72mm)を試作した。感度は、昨年度開発した二次元検出システムの数分の1であるが、露光と読み取りを同時に行うことができるので、蛋白質結晶構造解析実験などにおいて、X線シャッターなしで結晶を連続回転しながら自動測定を行えることなど、従来の二次元検出器にはない新しい機能を持った検出システムであることを示した。

Research Products
(27 results)

All Other

All Publications (27 results)

[Publications] 佐藤衛: "中性子で見るタンパク質の立体構造" 化学. 53・8. 64-65 (1998)
[Publications] M.Tomita: "Two Concentric Protein Shell Structure with Spikes of Silkworm Bombyx mori Cytoplasmic Polyhedrosis Virus Revealed by Small-angle Neutron Scattering Using the Contrast Variation Method" J.Biochem.(印刷中). (1999)
[Publications] A.Yamagauchi: "Characterization of Soluble Artificial Proteins with Random Sequences" FEBS Lett.421. 147-151 (1998)
[Publications] Y.Kamatari: "The Methanol-Induced Transition and the Expanded Helical Conformation in Hen Lysozyme" Protein Sci.7. 681-688 (1998)
[Publications] Y.Hagihara: "Chain-Like Conformation of Heat-Denatured Ribonuclease A and Cytochrome c as Evidenced by Solution X-ray Scattering" Folding & Design. 3. 195-201 (1998)
[Publications] J.C.Smith: "Motions in native and denatured proteins" physica B. 241-243. 1110-1114 (1998)
[Publications] T.Oka: "Conformational Change of Helix G in the Bacteriorhodopsin Photocycle : Investigation with Heavy Atom Labeling and X-ray Diffraction" Biophys.J.76. 1018-1023 (1999)
[Publications] S.Nomura: "Ordered Structure Formation of Bacteriorhodopsin-h DHFR in a Plasma Membrane" Langmuir. 15. 214-220 (1999)
[Publications] M.Kataoka: "Low Energy Dynamics of Globular Proteins Studied by Inelastic Neutron Scattering" J.Phys.Chem.Solid. (印刷中). (1999)
[Publications] Y.Kamatari: "The Compact and Expanded Penatured Conformations of Apomyoglobin in the Methanol-Water Solvent" Protein Sci.(印刷中). (1999)
[Publications] Y.Hasegawa: "Transmission-type X-ray linear polarizer with perfect crystals" J.Synchrotron Rad.5. 738-740 (1998)
[Publications] K.Okitsu: "X-ray triple refraction and triple absorption in a cobalt-complex crystal" J.Synchrotron Rad.5. 1055-1057 (1998)
[Publications] K.Okitsu: "X-ray linear birefringence and linear dichroism in a cobalt crystal measured with a tunable X-ray polarimeter" J.Synchrotron Rad.5. 995-997 (1998)
[Publications] H.Okuda: "Simultaneous dissolution and coarsening of δ precipitates in Al-Li alloys during DSC experiments" Zeitschrift fur METALLKUNDE. 88. 612-616 (1997)
[Publications] H.Okuda: "In-situ and Sumultaneous Synchrotron-Radiation Small-Angle and 100 Scattering Experiments on the Low-Temperature Structure in As-Quenched Al-Li Alloy during Heating" Scripta Materialia. 37. 1739-1744 (1997)
[Publications] H.Okuda: "Simultaneous SAS and experiments on phase decomposition and reversion in Al-Li binary alloys" Journal of Applied Cryst.30. 586-591 (1997)
[Publications] H.Okuda: "Nature of an endothermic peak of as-quenched Al-11.8mol% Li alloys" Materials Transactions. 39. 62-68 (1998)
[Publications] A.Minato: "Thermal and Structural Properties of sn-1,3-Dipalmitoyl-2-oleoylglycerol and sn-1,3-Dioleoyl-2-palmitoylglycerol Binary Mixtures Examined with Synchrotron Radiation X-ray Diffraction" J.Am.Oil Chem.Soc.74. 1213-1220 (1997)
[Publications] M.Arai: "Kinetic Refolding of β-Lactoglobulin. Studies by Synchrotron X-ray Scattering,and Circular Dichroism,Absorption and Fluorescence Spectroscopy" Journal of Molecular Biology. 275. 149-162 (1998)
[Publications] V.J.James: "Intermediate Filament Packing in α-Keratin of Echidna Quill" Textile Reserach Journal. 68. 167-170 (1998)
[Publications] V.J.James: "Molecular Structure Changes in Human Fetal Tissue during the Early Stages of Embryogenesis" Biochimica et Biophysica Acta. 1379. 282-288 (1998)
[Publications] Yu.A.Gaponov: "Object library for a new generation of experiment-controlling applications under the UNIX operating system" Journal of Synchrotron Rad.5. 593-595 (1998)
[Publications] Yu.A.Gaponov: "Structure changes in the sol-gel systems of hydrated oxides" Journal of Synchrotron Rad.5. 962-963 (1998)
[Publications] M.Koike: "Ti/Almultilayer zone plate and Bragg-Fresnel lens" Journal of Synchrotron Rad.5. 794-796 (1998)
[Publications] K.Wakabayashi: "High-resolution x-ray diffraction of muscle using undulator radiation from the Tristan main ring at KEK" Journal of Synchrotron Rad.5. 280-285 (1998)
[Publications] 佐藤衛: "タンパク質のX線解析" 共立出版, 123 (1998)
[Publications] 佐藤衛: "タンパク実験プロトコール(構造解析編)" 秀潤社, 260 (1997)

1998 Fiscal Year Annual Research Report

タンパク質の動的構造研究のためのX線溶液散乱解析システムの開発

Principal Investigator

佐藤 衛 横浜市立大学, 大学院・総合理学研究科, 教授 (60170784)

Research Products

[Publications] 佐藤 衛: "中性子で見るタンパク質の立体構造" 化学. 53・8. 64-65 (1998)

[Publications] M.Tomita: "Two Concentric Protein Shell Structure with Spikes of Silkworm Bombyx mori Cytoplasmic Polyhedrosis Virus Revealed by Small-angle Neutron Scattering Using the Contrast Variation Method" J.Biochem.(印刷中). (1999)

[Publications] A.Yamagauchi: "Characterization of Soluble Artificial Proteins with Random Sequences" FEBS Lett.421. 147-151 (1998)

[Publications] Y.Kamatari: "The Methanol-Induced Transition and the Expanded Helical Conformation in Hen Lysozyme" Protein Sci.7. 681-688 (1998)

[Publications] Y.Hagihara: "Chain-Like Conformation of Heat-Denatured Ribonuclease A and Cytochrome c as Evidenced by Solution X-ray Scattering" Folding & Design. 3. 195-201 (1998)

[Publications] J.C.Smith: "Motions in native and denatured proteins" physica B. 241-243. 1110-1114 (1998)

[Publications] T.Oka: "Conformational Change of Helix G in the Bacteriorhodopsin Photocycle : Investigation with Heavy Atom Labeling and X-ray Diffraction" Biophys.J.76. 1018-1023 (1999)

[Publications] S.Nomura: "Ordered Structure Formation of Bacteriorhodopsin-h DHFR in a Plasma Membrane" Langmuir. 15. 214-220 (1999)

[Publications] M.Kataoka: "Low Energy Dynamics of Globular Proteins Studied by Inelastic Neutron Scattering" J.Phys.Chem.Solid. (印刷中). (1999)

[Publications] Y.Kamatari: "The Compact and Expanded Penatured Conformations of Apomyoglobin in the Methanol-Water Solvent" Protein Sci.(印刷中). (1999)

[Publications] Y.Hasegawa: "Transmission-type X-ray linear polarizer with perfect crystals" J.Synchrotron Rad.5. 738-740 (1998)

[Publications] K.Okitsu: "X-ray triple refraction and triple absorption in a cobalt-complex crystal" J.Synchrotron Rad.5. 1055-1057 (1998)

[Publications] K.Okitsu: "X-ray linear birefringence and linear dichroism in a cobalt crystal measured with a tunable X-ray polarimeter" J.Synchrotron Rad.5. 995-997 (1998)

[Publications] H.Okuda: "Simultaneous dissolution and coarsening of δ precipitates in Al-Li alloys during DSC experiments" Zeitschrift fur METALLKUNDE. 88. 612-616 (1997)

[Publications] H.Okuda: "In-situ and Sumultaneous Synchrotron-Radiation Small-Angle and 100 Scattering Experiments on the Low-Temperature Structure in As-Quenched Al-Li Alloy during Heating" Scripta Materialia. 37. 1739-1744 (1997)

[Publications] H.Okuda: "Simultaneous SAS and experiments on phase decomposition and reversion in Al-Li binary alloys" Journal of Applied Cryst.30. 586-591 (1997)

[Publications] H.Okuda: "Nature of an endothermic peak of as-quenched Al-11.8mol% Li alloys" Materials Transactions. 39. 62-68 (1998)

[Publications] A.Minato: "Thermal and Structural Properties of sn-1,3-Dipalmitoyl-2-oleoylglycerol and sn-1,3-Dioleoyl-2-palmitoylglycerol Binary Mixtures Examined with Synchrotron Radiation X-ray Diffraction" J.Am.Oil Chem.Soc.74. 1213-1220 (1997)

[Publications] M.Arai: "Kinetic Refolding of β-Lactoglobulin. Studies by Synchrotron X-ray Scattering,and Circular Dichroism,Absorption and Fluorescence Spectroscopy" Journal of Molecular Biology. 275. 149-162 (1998)

[Publications] V.J.James: "Intermediate Filament Packing in α-Keratin of Echidna Quill" Textile Reserach Journal. 68. 167-170 (1998)

[Publications] V.J.James: "Molecular Structure Changes in Human Fetal Tissue during the Early Stages of Embryogenesis" Biochimica et Biophysica Acta. 1379. 282-288 (1998)

[Publications] Yu.A.Gaponov: "Object library for a new generation of experiment-controlling applications under the UNIX operating system" Journal of Synchrotron Rad.5. 593-595 (1998)

[Publications] Yu.A.Gaponov: "Structure changes in the sol-gel systems of hydrated oxides" Journal of Synchrotron Rad.5. 962-963 (1998)

[Publications] M.Koike: "Ti/Almultilayer zone plate and Bragg-Fresnel lens" Journal of Synchrotron Rad.5. 794-796 (1998)

[Publications] K.Wakabayashi: "High-resolution x-ray diffraction of muscle using undulator radiation from the Tristan main ring at KEK" Journal of Synchrotron Rad.5. 280-285 (1998)

[Publications] 佐藤 衛: "タンパク質のX線解析" 共立出版, 123 (1998)

[Publications] 佐藤 衛: "タンパク実験プロトコール(構造解析編)" 秀潤社, 260 (1997)

佐藤衛横浜市立大学, 大学院・総合理学研究科, 教授 (60170784)

[Publications] 佐藤衛: "中性子で見るタンパク質の立体構造" 化学. 53・8. 64-65 (1998)

[Publications] 佐藤衛: "タンパク質のX線解析" 共立出版, 123 (1998)

[Publications] 佐藤衛: "タンパク実験プロトコール(構造解析編)" 秀潤社, 260 (1997)