1997 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) 片岡幹雄大阪大学, 大学院・理学研究科, 助教授 (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、動的構造研究に用いる試料槽を開発することを目的に、各種スペクトル測定用に市販されているストップトフロー混合装置を改良して、X線溶液散乱測定装置に組み込んだ。改良された本装置の特徴は、(1)試料溶液温度を-20℃から95℃まで0.1℃以内に制御でき、かつ測定中に温度をモニターしてその変動を記録できる、(2)試料を注入するキャピラリーの脱着を容易にして、バックグラウンドの補正が簡単にできる、(3)高圧下での測定も可能である、ことなどで、本装置の利用により様々な条件でのX線溶液散乱実験が可能になるものと期待される。一方、片岡は、データ解析システムの開発の一環として、X線溶液散乱法の最大の利点の一つである非天然状態の溶液構造情報を得るための解析法を開発し、その手法をαラクトアルブミンの変性過程の解析に応用した。その結果、αラクトアルブミンのモルテングロビュール状態の構造を詳細に特徴づけ、他の球状タンパク質のMGの構造と比較し、MGの多様性を明らかにした。また、雨宮は、X線イメージインテンシファイヤーと冷却CCDを組み合わせた高感度2次元X線検出器の開発を行い、所期の性能を定量的に評価した。その結果、(1)感度は量子検出効率で約60%であり、イメージングプレートに比べて2倍の高感度であること、(2)X線イメージインテンシファイヤーのダイナミックレンジは6桁あるが、ファクター2の非線形性があることを見出すとともに、(3)画像歪みと感度の不均一性を補正するためのハードウエアとソフトウエアの開発を行い、補正により画像歪み、感度の不均一性、ともに1%程度が実現することに成功した。

Research Products
(23 results)

All Other

All Publications (23 results)

[Publications] 佐藤衛: "中性子による計測と利用-中性子小角散乱(生物)" Radioisotopes. 46. 193-200 (1997)
[Publications] Y.Nakamura: "Small-angle X-ray scattering with Imaging Plate.Application to dilute polymer Solutions" Polymer Bulletin. 38. 469-476 (1997)
[Publications] K.Okada: "Three-dimensional structure of Escherichia coli branched-chain amino acid aminotransferase at 2.5 A resolution" Journal of Biochemistry. 121. 637-641 (1997)
[Publications] M.Kataoka: "Structural characterization of molten globule of a-lactalbumin by solution X-ray Scattering" Protein Science. 6. 422-430 (1997)
[Publications] K.Mihara: "Functional expression and site-directed mutaganesis of photoactive yellow proteim" Journal of Biochemistry. 121. 876-880 (1997)
[Publications] Y.Imamoto: "Evidence for proton transfer from Glu-46 to the chrompophore during the photocycle of photoactive yellow protein" Journal of Biological Chemistry. 272. 12905-12908 (1997)
[Publications] L.S.Brown: "Global shift of protein structure triggered by local electrostatic change in bacteriorhodopsin" Proceedings of National Academy of Science U.S.A.94. 5040-5044 (1997)
[Publications] T.Konno: "Urea-induced denatured states of a protein,Streptomyces subtilisin inhibitor" Protein Science. 6. 2242-2249 (1997)
[Publications] T.Konno: "A circular dichroism study of preferential hydration and alcohol effects on a denatured protein,pig calpastatin domain I" Biochimica et Biophysica Acta. 1342. 73-82 (1997)
[Publications] H.Kamikubo: "The last phase of the reprotonation switch in bacteriorhodopsin : The transition between the M-type and the N-type protein conformation depends on hydration" Biochemistry. 36. 12282-12287 (1997)
[Publications] T.Oka: "X-ray diffraction studies of bacteriorhodopsin:Determination of the positions of mercury label at several engineered cysteine residues" Photochemistry and Photobiology. 66. 768-773 (1997)
[Publications] A.V.Goupil-Lamy: "High-resolution vibrational spectroscopy of a globular protein:Inelastic neutron Scattering and normal mode analysis of staphylococcal nuclease" Journal of American Chemical Society. 119. 9268-9272 (1997)
[Publications] M.Hoshino: "Trifluoroethanol-induced conformational transition of hen egg-white lysozyme studied by small-angle X-ray scattering" FEBS Letters. 416. 72-76 (1997)
[Publications] K.Inaba: "Structural and functional roles of modules in hemoglobin-Substitution of module M4 in hemoglobin subunits" Journal of Biological Chemistry. 272. 30054-30060 (1997)
[Publications] Y.O.Kamatari: "The methanol-induced transition and the expanded helical conformation of lysozyme" Protein Science. (in press).
[Publications] J.C.Smith: "Motions in Native and Denatured Proteins" Physica B. (in press).
[Publications] A.Minato: "Thermodynamic and kinetic study on phase behavior of binary mixtures of POP and PPO forming molecular compound systems" J.Phys.Chem.B. 101. 3498-3505 (1997)
[Publications] S.Ueno: "Synchrotron radiation x-ray diffraction study of liquid crystal formation and polymorphic crystallization of SOS" J.Phys.Chem.B. 101. 6847-6854 (1997)
[Publications] M.F.Mina: "Dymanic investigation of the solid-solid phase transition of normal-alkane by simultaneous measurement with differential scanning calorimetry,small-angle x-ray scattering and x-ray television detector" Jpn.J.Appl.Phys.36. 5616-5622 (1997)
[Publications] J.M.Zhou: "Unfolding of dimeric creatine kinase in urea and guanidine hydrochloride as measured using small-angle x-ray scattering with synchrotron radiation" FEBS Letters. 415. 183-185 (1997)
[Publications] 佐藤衛: "構造生物学とその解析法" 共立出版, 192 (1997)
[Publications] 片岡幹雄: "蛋白質のかたちと物性" 共立出版, 248 (1997)
[Publications] Y.Amemiya: "Methods in Ehzymology,Vol.276" Academic Press, Inc., 700 (1997)

1997 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: "中性子による計測と利用-中性子小角散乱(生物)" Radioisotopes. 46. 193-200 (1997)

[Publications] Y.Nakamura: "Small-angle X-ray scattering with Imaging Plate.Application to dilute polymer Solutions" Polymer Bulletin. 38. 469-476 (1997)

[Publications] K.Okada: "Three-dimensional structure of Escherichia coli branched-chain amino acid aminotransferase at 2.5 A resolution" Journal of Biochemistry. 121. 637-641 (1997)

[Publications] M.Kataoka: "Structural characterization of molten globule of a-lactalbumin by solution X-ray Scattering" Protein Science. 6. 422-430 (1997)

[Publications] K.Mihara: "Functional expression and site-directed mutaganesis of photoactive yellow proteim" Journal of Biochemistry. 121. 876-880 (1997)

[Publications] Y.Imamoto: "Evidence for proton transfer from Glu-46 to the chrompophore during the photocycle of photoactive yellow protein" Journal of Biological Chemistry. 272. 12905-12908 (1997)

[Publications] L.S.Brown: "Global shift of protein structure triggered by local electrostatic change in bacteriorhodopsin" Proceedings of National Academy of Science U.S.A.94. 5040-5044 (1997)

[Publications] T.Konno: "Urea-induced denatured states of a protein,Streptomyces subtilisin inhibitor" Protein Science. 6. 2242-2249 (1997)

[Publications] T.Konno: "A circular dichroism study of preferential hydration and alcohol effects on a denatured protein,pig calpastatin domain I" Biochimica et Biophysica Acta. 1342. 73-82 (1997)

[Publications] H.Kamikubo: "The last phase of the reprotonation switch in bacteriorhodopsin : The transition between the M-type and the N-type protein conformation depends on hydration" Biochemistry. 36. 12282-12287 (1997)

[Publications] T.Oka: "X-ray diffraction studies of bacteriorhodopsin:Determination of the positions of mercury label at several engineered cysteine residues" Photochemistry and Photobiology. 66. 768-773 (1997)

[Publications] A.V.Goupil-Lamy: "High-resolution vibrational spectroscopy of a globular protein:Inelastic neutron Scattering and normal mode analysis of staphylococcal nuclease" Journal of American Chemical Society. 119. 9268-9272 (1997)

[Publications] M.Hoshino: "Trifluoroethanol-induced conformational transition of hen egg-white lysozyme studied by small-angle X-ray scattering" FEBS Letters. 416. 72-76 (1997)

[Publications] K.Inaba: "Structural and functional roles of modules in hemoglobin-Substitution of module M4 in hemoglobin subunits" Journal of Biological Chemistry. 272. 30054-30060 (1997)

[Publications] Y.O.Kamatari: "The methanol-induced transition and the expanded helical conformation of lysozyme" Protein Science. (in press).

[Publications] J.C.Smith: "Motions in Native and Denatured Proteins" Physica B. (in press).

[Publications] A.Minato: "Thermodynamic and kinetic study on phase behavior of binary mixtures of POP and PPO forming molecular compound systems" J.Phys.Chem.B. 101. 3498-3505 (1997)

[Publications] S.Ueno: "Synchrotron radiation x-ray diffraction study of liquid crystal formation and polymorphic crystallization of SOS" J.Phys.Chem.B. 101. 6847-6854 (1997)

[Publications] M.F.Mina: "Dymanic investigation of the solid-solid phase transition of normal-alkane by simultaneous measurement with differential scanning calorimetry,small-angle x-ray scattering and x-ray television detector" Jpn.J.Appl.Phys.36. 5616-5622 (1997)

[Publications] J.M.Zhou: "Unfolding of dimeric creatine kinase in urea and guanidine hydrochloride as measured using small-angle x-ray scattering with synchrotron radiation" FEBS Letters. 415. 183-185 (1997)

[Publications] 佐藤 衛: "構造生物学とその解析法" 共立出版, 192 (1997)

[Publications] 片岡幹雄: "蛋白質のかたちと物性" 共立出版, 248 (1997)

[Publications] Y.Amemiya: "Methods in Ehzymology,Vol.276" Academic Press, Inc., 700 (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: "中性子による計測と利用-中性子小角散乱(生物)" Radioisotopes. 46. 193-200 (1997)

[Publications] 佐藤衛: "構造生物学とその解析法" 共立出版, 192 (1997)