1995 Fiscal Year Annual Research Report

蛋白質の機能構造研究のためのX線結晶解析法の迅速高精度化

Research Project

Project/Area Number	06305006
Research Institution	University of Tokyo
Principal Investigator	佐藤能雅東京大学, 薬学部, 教授 (30150014)
Co-Investigator(Kenkyū-buntansha)	福山恵一大阪大学, 理学部, 教授 (80032283) 月原冨武大阪大学, 蛋白質研究所, 教授 (00032277) 三木邦夫京都大学, 大学院理学研究科, 教授 (10116105) 田中勲北海道大学, 理学部, 教授 (70093052) 田中信夫東京工業大学, 生命理工学部, 教授 (50032024)
Keywords	蛋白質 / X線結晶構造解析 / 膜蛋白質 / 糖蛋白質 / 抗体 / 結晶化 / 三次元構造 / 構造生物学
Research Abstract	蛋白質の機能の解明と応用の研究を進展させるため、蛋白質の三次元構造の研究の分担を通じ、X線結晶解析の方法の新展開と、蛋白質の機能構造の迅速な研究体制の構築を本研究の目的とした。平成7年度では、1 アミノ酸改変蛋白質、2 各種プロテアーゼとその阻害剤との複合体、3 アミラーゼ等とその阻害蛋白質、4 核酸および核酸を認識する蛋白質、5 抗原を認識する抗体と抗原複合体、6 膜蛋白質結晶化法、7 ウイルス等の巨大分子単位、その直接解析法、8 アミノ酸改変蛋白質の産製と迅速解析法、9 高速高精度なX線回析データ測定法、10 X線異常分散を利用する解析法、11 迅速な自動分子置換構造解析システムの構築、12 糖蛋白質の結晶化と解析の手法、13 溶液NMR情報を用いるX線解析法など、蛋白質構造の解析と、新解析手法の開発と構築に関する研究を行い、これらに関する成果を得た。研究の成果として、系統化した結晶化法と結晶化用の試料調製法に基づいて、膜蛋白質bovine cytochrome c oxidaseと糖蛋白質human renal dipeptidaseの解析に成功した。高精度データ測定の装置システムとソフトウェアCa等の異常分散効果の利用法、多波長異常分散法による直接解析法、自動分子置換解析法などを開発・構築したキメラ酵素の作成とその構造の解析、酵素の耐熱化の機構の研究、トリプシン族の酵素とその阻害剤との複合体の解析、解糖系等の酵素およびその阻害蛋白質の解析、抗体のFvとFabのフラグメントと抗原ハプテンとの複合体の解析などを行った。方法論的な展開と研究者間の連携と協力をはかる本研究の骨子を達成するため、公開の研究会を開催し、さらに、研究の進捗と成果を取りまとめた研究成果報告書を印刷し、配布した。

Research Products

(19 results)

All Other

All Publications (19 results)

[Publications] Mizutani, Ryuuta 他: "Three-dimesional Structures of the Fab Fragment of Murine NIG9 Antibody from the Primary Immune Response and of Its Complex with (4-Hydroxy-3-nitrophenyl) acetate." J. Mol. Biol.254. 208-222 (1995)
[Publications] Noguchi, shuji 他: "Crystal Structure of Ustilago sphaerogena Ribonuclease U_2 at 1.8 Å Resolution." Biochemistry. 34. 15583-15591 (1995)
[Publications] Harada, Shigeharu 他: "Crystallization and Preliminary X-ray Diffraction Studies of Curculin, A New Type of Sweet Protein Having Taste-Midifying Action." J. Mol. Biol.238. 286-287 (1994)
[Publications] Harada, Shigeharu 他: "Complete Amino Acid Sequence of a Zinc Metalloendoprotease from Streptomyces caespitosus." Eur. J. Biochem.233. 683-686 (1995)
[Publications] Nakamura, Msahiro 他: "Design of Noncovalent Trypsin Inhibitor Based on the X-ray Crystal Structure of the Complex." Biochem. Biopys. Res. Comm.213. 583-587 (1995)
[Publications] Takenaka, Akio 他: "Structural Composition of Hammerhead Ribozymes." J. Biochem.119. 49-57 (1996)
[Publications] Kumasaka, Takashi 他: "Crystal Structures of H_2C Proteinase from the Venom of Trimeresurus flavoviridis." J. Biochem.119. 49-57 (1996)
[Publications] Nagata, Chikahiro 他: "Crystallization and Preliminary X-ray Analysis of Cytochrome c_<552> from Pseudomonas europae." J. Biochem.116. 946-947 (1995)
[Publications] Tsukihara, Tomitake 他: "Structures of Metal Sites of Oxidized Bovine Cytochrome c Oxidase at 2.8 Å." Science. 269. 1069-1074 (1995)
[Publications] Gohda, Keigo 他: "Crystal Structure of RNase T_1 Complexed with 3'AMP: Structural Evidence for Direct Interaction of His 40 and Glu 58 with the 2'-Hydroxyl Group of the Ribose." J. Biol. Chem.269. 17531-17536 (1994)
[Publications] Miyatake, Hideyuki 他: "Crystal Structure of the Unliganded Alkaline Protease from Pseudomonas aeruginosa IFO3080 and Its Conformational Changes on Ligand Binding." J. Biochem.118. 474-479 (1995)
[Publications] Takano, Kazufumi 他: "Contribution of Hydrophobic Residues to the Stability of Human Lysozyme: Calorimetric Studies and X-ray Structural Analysis of the Five Isoleucine to Valine Mutants." J. Mol. Biol.254. 62-76 (1995)
[Publications] Yamane, Takashi 他: "Crystal Structure of Streptomyces erythraeus Trypsin at 1.9 Å Resolution." J. Biochem.118. 882-894 (1995)
[Publications] Nitanai, Yasushi 他: "Crystallizationand Preliminary X-Ray Investigation of a Glycoprotein, Human Renal Dipeptidase." J. Crystal Growth.印刷中. (1996)
[Publications] Miki, Kunio 他: "Application of Automatic Molecular Replacement Procedure to Crystal Structure Analysis of Cytochrome c_2 from Rhodopseudomonas viridis." Acta Crystallogr.D50. 271-275 (1994)
[Publications] Suzuki, Atsuo 他: "Crystal Structure Analysis of Phospholipase A2 from Trimeresurus flavoviridis (Habu Snake) at 1.5 Å Resolution." J. Biochem.117. 730-740 (1995)
[Publications] Yamagata, Yuriko 他: "Medel Building of DNA Repair Enzyme 3-Methyladenosine DNA Glycosidase-DNA Complex." Nucleic Acids Symposium Series. 34. 7-8 (1995)
[Publications] Mizuyani, Ryuta 他: "Crystallization and Crystallographic Studies on Fab Fragments of Anti-Nitrophenol Antibodies from Primary and Secondary Immune Responses." J. Crystal Growth. 印刷中. (1996)
[Publications] 佐藤能雅: "“続医薬品の開発"第17巻「薬物の構造解析」(荒田洋治編集のp. 66-97を担当)" 廣川書店, 222 (1995)

1995 Fiscal Year Annual Research Report

蛋白質の機能構造研究のためのX線結晶解析法の迅速高精度化

Principal Investigator

佐藤 能雅 東京大学, 薬学部, 教授 (30150014)

Research Products

[Publications] Mizutani, Ryuuta 他: "Three-dimesional Structures of the Fab Fragment of Murine NIG9 Antibody from the Primary Immune Response and of Its Complex with (4-Hydroxy-3-nitrophenyl) acetate." J. Mol. Biol.254. 208-222 (1995)

[Publications] Noguchi, shuji 他: "Crystal Structure of Ustilago sphaerogena Ribonuclease U_2 at 1.8 Å Resolution." Biochemistry. 34. 15583-15591 (1995)

[Publications] Harada, Shigeharu 他: "Crystallization and Preliminary X-ray Diffraction Studies of Curculin, A New Type of Sweet Protein Having Taste-Midifying Action." J. Mol. Biol.238. 286-287 (1994)

[Publications] Harada, Shigeharu 他: "Complete Amino Acid Sequence of a Zinc Metalloendoprotease from Streptomyces caespitosus." Eur. J. Biochem.233. 683-686 (1995)

[Publications] Nakamura, Msahiro 他: "Design of Noncovalent Trypsin Inhibitor Based on the X-ray Crystal Structure of the Complex." Biochem. Biopys. Res. Comm.213. 583-587 (1995)

[Publications] Takenaka, Akio 他: "Structural Composition of Hammerhead Ribozymes." J. Biochem.119. 49-57 (1996)

[Publications] Kumasaka, Takashi 他: "Crystal Structures of H_2C Proteinase from the Venom of Trimeresurus flavoviridis." J. Biochem.119. 49-57 (1996)

[Publications] Nagata, Chikahiro 他: "Crystallization and Preliminary X-ray Analysis of Cytochrome c_<552> from Pseudomonas europae." J. Biochem.116. 946-947 (1995)

[Publications] Tsukihara, Tomitake 他: "Structures of Metal Sites of Oxidized Bovine Cytochrome c Oxidase at 2.8 Å." Science. 269. 1069-1074 (1995)

[Publications] Gohda, Keigo 他: "Crystal Structure of RNase T_1 Complexed with 3'AMP: Structural Evidence for Direct Interaction of His 40 and Glu 58 with the 2'-Hydroxyl Group of the Ribose." J. Biol. Chem.269. 17531-17536 (1994)

[Publications] Miyatake, Hideyuki 他: "Crystal Structure of the Unliganded Alkaline Protease from Pseudomonas aeruginosa IFO3080 and Its Conformational Changes on Ligand Binding." J. Biochem.118. 474-479 (1995)

[Publications] Takano, Kazufumi 他: "Contribution of Hydrophobic Residues to the Stability of Human Lysozyme: Calorimetric Studies and X-ray Structural Analysis of the Five Isoleucine to Valine Mutants." J. Mol. Biol.254. 62-76 (1995)

[Publications] Yamane, Takashi 他: "Crystal Structure of Streptomyces erythraeus Trypsin at 1.9 Å Resolution." J. Biochem.118. 882-894 (1995)

[Publications] Nitanai, Yasushi 他: "Crystallizationand Preliminary X-Ray Investigation of a Glycoprotein, Human Renal Dipeptidase." J. Crystal Growth.印刷中. (1996)

[Publications] Miki, Kunio 他: "Application of Automatic Molecular Replacement Procedure to Crystal Structure Analysis of Cytochrome c_2 from Rhodopseudomonas viridis." Acta Crystallogr.D50. 271-275 (1994)

[Publications] Suzuki, Atsuo 他: "Crystal Structure Analysis of Phospholipase A2 from Trimeresurus flavoviridis (Habu Snake) at 1.5 Å Resolution." J. Biochem.117. 730-740 (1995)

[Publications] Yamagata, Yuriko 他: "Medel Building of DNA Repair Enzyme 3-Methyladenosine DNA Glycosidase-DNA Complex." Nucleic Acids Symposium Series. 34. 7-8 (1995)

[Publications] Mizuyani, Ryuta 他: "Crystallization and Crystallographic Studies on Fab Fragments of Anti-Nitrophenol Antibodies from Primary and Secondary Immune Responses." J. Crystal Growth. 印刷中. (1996)

[Publications] 佐藤能雅: "“続医薬品の開発"第17巻「薬物の構造解析」(荒田洋治編集のp. 66-97を担当)" 廣川書店, 222 (1995)

佐藤能雅東京大学, 薬学部, 教授 (30150014)