• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

時間分割蛋白質結晶構造解析法の研究

Research Project

Project/Area Number 05244102
Research Category

Grant-in-Aid for Scientific Research on Priority Areas

Allocation TypeSingle-year Grants
Research InstitutionNagoya University

Principal Investigator

山根 隆 (1996)  名古屋大学, 工学部, 教授 (80030055)

芦田 玉一 (1993-1995)  名古屋大学, 工学部, 教授 (10029936)

Co-Investigator(Kenkyū-buntansha) 三木 邦夫  京都大学, 大学院・理学研究科, 教授 (10116105)
月原 富武 (月原 冨武)  大阪大学, 蛋白質研究所, 教授 (00032277)
田中 信夫  東京工業大学, 生命理工学部, 教授 (50032024)
佐藤 能雅 (佐藤 能稚)  東京大学, 薬学部, 教授 (30150014)
安岡 則武  姫路工業大学, 理学部, 教授 (40029054)
Project Period (FY) 1993 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥143,100,000 (Direct Cost: ¥143,100,000)
Fiscal Year 1996: ¥14,600,000 (Direct Cost: ¥14,600,000)
Fiscal Year 1995: ¥16,500,000 (Direct Cost: ¥16,500,000)
Fiscal Year 1994: ¥55,000,000 (Direct Cost: ¥55,000,000)
Fiscal Year 1993: ¥57,000,000 (Direct Cost: ¥57,000,000)
Keywords蛋白質結晶 / 動的解析 / ラウエ法 / 放射光 / 酵素反応トレース / 良質結晶作成法 / 温度ジャンプ / pHジャンプ / 蛋白質結晶構造解析 / 蛋白質分子構造 / 酵素反応機構 / 蛋白質結晶化 / X線結晶構造解析 / フローセル法 / 迅速解析 / 分子置換法 / 時間分割ラウエ法 / 酵素反応
Research Abstract

本重点領域研究では、蛋白質結晶学における通常の静的な構造解析を越える動的な解析を目指して、研究分担者が、以下の役割を担当した。得られた研究成果は、得られた研究成果は、当重点領域研究のNews Letterや公開シンポジウムで発表された。主要な研究実績は以下の通りである。山根:「酵素反応のトレース」で、「フローセルを用いたラウエ法の加水分解酵素への適用」で、M-プロテアーゼのアルカリ安定化機構を解明した。「モジュール置換キメラへモグロビンの結晶構造解析」で大希望に改変した蛋白質の構造と機能を解明した。安岡:「電子伝達反応の研究」でチトクロムc'のヘムに配位子を結合させ、ヘム周辺のアミノ酸側鎖の移動を観察し、機能の解明を進めた。佐藤:「酵素反応トレース」で「糖蛋白質ヒト腎臓ジペプチダーゼの低温結晶構造解析」を行い、活性発現機構を検討した。田中:赤外線ランプおよびレーザー用いる2種類の温度ジャンプ法の開発に成功した。「酵素耐熱化機構の研究」でレーザー照射前後でのイソプロピルリンゴ酸脱水素酵素の構造変化を観測し、耐熱性の検討を行った。月原:「良質結晶作成法の開発、分子置換法解析法の開発」で超分子複合体であるチトクロム酸化酵素の良質の結晶を調整し、2.8A分解能の結晶構造を世界で初めて解明した。三木:「良質結晶作成法の開発、結晶評価法の開発」で、結晶化が極めて困難な膜蛋白質である光合成反応中心複合体の結晶化など多数の蛋白質の結晶化に成功した。松浦:結晶化に用いる沈殿剤の役割と結晶成長との関係を検討した。箱島「実験室系でのX線強度測定系の構築」を行い、性能を評価した。相原:微小重力場では結晶成長過程に違いのあることを示した。井上:「電子伝達酵素反応のトレース」でプラストシアニンの動的解析のためにラウエ法データで構造解析を行い、単色光データと同程度の精密な分子の立体構造を決定した。

Report

(4 results)
  • 1996 Annual Research Report
  • 1995 Annual Research Report
  • 1994 Annual Research Report
  • 1993 Annual Research Report
  • Research Products

    (63 results)

All Other

All Publications (63 results)

  • [Publications] A.Suzuki,T.Yamane,T.Ashida: "Crystallographic refinement of Bowman-Birk type protease inhibitor A-II from Peanut (Arachis hypogaea) at 2.3A resolution." J.MOl.Biol.234. 722-734 (1993)

    • Related Report
      1996 Annual Research Report
  • [Publications] A.Suzuki,T.Yamane,T.Ashida: "Crystallization and preliminary X-ray studies on the trypsin inhibitor I-2 from wheat germ and its complex with trypsin." Acta Crystallogr.D49. 594-596 (1993)

    • Related Report
      1996 Annual Research Report
  • [Publications] T.Yamane,A.Suzuki,T.Ashida: "Crystal structure of a new aikaline serine protease (M-protease) from Bacillus sp.KSM-K16" Acta Crystallogr.D51. 199-206 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] A.Suzuki,T.Yamane,T.Ashida: "Crystal structure analysis of phospholipase A2 from Trimeresurus flavoviridis (Habu snake) venom at 1.5A resolution." J.Biochem.117. 730-740 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] T.Yamane,A.Suzuki,A.Ashida: "Crystal structure of Streptomyces erythraeus trypsin at 1.9A resolution." J.Biochem.118. 882-894 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] T.Shirai,A.Suzuki,T.Yamane,T.Ashida: "High-resolution crystal structure of M-protease : phylogeny aided analysis of the high-alkaline adaptation mechanism." Protein Engineering. (in press). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Y.Higuchi,N.Yasuoka,T.E.Meyer,M.A.Cusanovich: "Crystallization and preliminary X-ray study of a new crystal form of cytochrome c' from Rhodobacter capsulatus" Acta Cryst.D52. 199-200 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] T.H.Tahirov,S.Misakai,Y.Higuchi,N.Yasuoka: "High-resolution crystal structures of two polymorphs of cytochrome c' from purple phototrophic bacterium Rhodobacter capsulatus" J.Mol.Biol.259. 467-479 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] T.H.Tahirov,S.Misakai,T.E.Meyer,N.Yasuoka: "Concerned movement of side chains in the haem vicinity observed on ligand binding in cytochrome c' from Rhodobacter capsulatus" Nature Structural Biology. 3. 459-464 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] S.Noguchi,Y.Satow,C.Sasaki,T.Matsuzaki: "Crystal Stracture of Ustilago shpaerogena Ribonuclease U2 at 1.8Å Resulution" Biochemistry. 34. 15583-15591 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] S.Noguchi,Y.Satow,C.Sasaki,T.Matsuzaki: "Crystallization and preliminary X-ray investigatio of six new crystal forms of ribonuclease U2 from Ustilago sphaerogena" Journal of Crystal Growth. 168. 270-274 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Y.Nitanai,Y.Satow,H.Adachi,M.Tsujimoto: "Cyrstallization and preliminary X-ray investigation of a glycoprotein,human renal dipeptidase" Journal of Crystal Growth. 168. 280-283 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] M.Nishida,S.Harada,Y.Satow,K.Takahashi: "Crystallization and X-ray diffraction studies of glutathione S-transferase from Escherichia coli" Journal of Crystal Growth. 168. 284-287 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] K.Miyawaki,S.Noguchi,S.Harada,Y.Satow: "Crystallography of succinimide hen egg-white lysozyme at low temperatures" Journal of Crystal Growth. 168. 292-296 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] M.Sakurai,M.Ohzeki,N.Tanaka,T.Oshima: "Structure of a Loop-Deleted Variant of 3-Isopropylmalate Dehydrogenase from Thermus thermophilus : an Internal Reprieve Tolerance Mechanism" Acta Cryst..D52. 124-128 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] C.Nagata,H.Moriyama,N.Tanaka,T.Oshima: "Crystallography of 3-Isopropylmalate Dehydrogenase from Thermus the rmophillus and its Chimeric Enzyme" Acta Cryst..D52. 623-630 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] N.Igarashi,H.Moriyama,T.Mikami and N.Tanaka: "Detwinning of hemihedrally twinned crystals by the least squares method and its application to the crystal of hydroxylamine oxidoreductase from Nitrosomonas europaea" To be pubished.

    • Related Report
      1996 Annual Research Report
  • [Publications] T.Tsukihara,H.Aoyama,R.Yaono,S.Yoshikawa: "The Whole Structure of the 13-Subunit Oxidized Cytochrome c Oxidase at 2.8Å" Science. 272. 1136-1144 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] K.Miki,S.Sogabe,A.Uno,T.Ezoe,N.Kasai: "Application of automatic molecular replacement procedure to crystal structure analysis of cytochrome c2 from R.viridis" Acta Crystallogr.D50. 271-275 (1994)

    • Related Report
      1996 Annual Research Report
  • [Publications] S.Sogabe,T.Ezoe,N.Kasai,M.Saeda & K.Miki: "Structural similarity of cytochrome c2 from R.viridis to mitochondrial cytochrome c reversed by its crystal structure" FEBS Lett.345. 5-8 (1994)

    • Related Report
      1996 Annual Research Report
  • [Publications] N.Katayama,M.Kobayasi F.Motojima,K.Miki: "Preliminary X-ray crystallographic studies of photosynthetic reaction center from a thermophilic sulfur bacterium,C.tepidum" FEBS Lett.348. 158-160 (1994)

    • Related Report
      1996 Annual Research Report
  • [Publications] K.Saika,K.Inaka,T.Matsui,K.Miki: "Crystallization of mutant β subunit of F1-ATPase from thermophilic Bacillus PS3" J.Mol.Biol.242. 709-711 (1994)

    • Related Report
      1996 Annual Research Report
  • [Publications] A.Kita,S.Kasai,K.Miki: "Crystal structure determination of a flavoprotein FP390 from a luminescent bacterium,P.phosphoreum" J.Biochem.117. 575-578 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] H.Nishida,K.Inaka,K.Miki: "Crystal structure of NADH-cytochrome b5 reductase from pig liver at 2.4A resolution" Biochemistry. 34. 2763-2767 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] H.Nishida,K.Inaka & K.Miki: "Specific arragement of three amino acid residues for flavin-binding barrel structures in NADH-cytochrome b5 reductase" FEBS Lett.361. 97-100 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] T.Tamada,H.Nishida,K.Miki: "A new crystal form of photolyase (photoreactivating enzyme) from the cyanobacterium A.nidulans" J.Struct.Biol.115. 37-40 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] K.Inaka,M.Kikuchi,K.Miki: "Crystal structure of a gultathionylated mutant human lysozyme : A folding intermediate mimic in the formation of a disulfide bond" Acta Crystallogr.D51. 619-625 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] J.Saito,Y.Nagata,K.Miki: "Crystallization and preliminary X-ray crystallographic analysis of chitosanase from B.circulans MH-K1" Acta Crystallogr.D51. 856-857 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] S.Sogabe & K.Miki: "Refined crystal structure of ferrocytochrome c2 from R.viridis at 1.6A resolution" J.Mol.Biol.252. 235-247 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] F.Motojima,K.Inaka,K.Miki: "Preliminary X-ray diffraction studies of a new crystal form of C-1027-AG,the apoprotein of the macromolecular antitumor antibio" Acta Crystallor.D51. 1084-1085 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] A.Kita,S.Kasai,M.Miyata & K.Miki: "The structure of flavoprotein FP390 from luminescent bacterium,P.phosphoreum at 2.7A resolution" Acta Crystallogr.D52. 77-86 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] M.Kataoka,K.Yamamoto K.Miki: "Crystallization and preliminary X-ray diffraction study of aldehyde reductase from a red yeast,S.salmonicolor" Acta Crystallogr.D52. 405-406 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] H.Nishida & K.Miki: "Electrostatic properties deduced from refined structures of NADH-cytochrome b5 reductase and the other flavin-dependent reductases" Protein : Structure,Function and Genetics. 26. 32-41 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] M.Sugahara,N.Sekine-Suzuki,K.Miki: "Crystallization and preliminary X-ray diffraction studies of toxin (perfringolysin O),a pore-forming cytolysin of C.perfringens" J.Crystal Growth. 168. 288-291 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] T.A.Funaki,Y.Takasuga,K.Miki: "Crystallization and preliminary X-ray characterization of Chaperonin 60 from P.denitrificans" J.Crystal Growth. 168. 297-300 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] M.Tsunemi: "Crystal Structure of an Elastase-Specific Inhibitor Elafin Complexed with Porcine Pancreatic Elastase Determined at 1.9A resolution." Biochemistry. 35. 11570-11576 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Y.Morishita: "Crystal Structure of a Maltotetraose-forming Exo-amylase from Pseudomonas stutzeri." J.Mol.Biol.267. 1-12 (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] T.Inoue: "Crystallization and Preliminary X-Ray Studies on Pseudoazurin from Achromobacter cycloclates IAM1013" J.Biochem. 114. 761-762 (1993)

    • Related Report
      1996 Annual Research Report
  • [Publications] T.Inoue: "Refined Crystal Structure of Pseudoazurin from Methylobacterium extorquens AM1 at 1.5A Resolution." Acta Cryst.D50(3). 317-328 (1994)

    • Related Report
      1996 Annual Research Report
  • [Publications] T.Inoue: "Three-Dimensional Structure of Azurin from Achromobacter xylosoxidas NCIB11015 at 2.5A Resolution." J.Biochem.116. 1193-1197 (1994)

    • Related Report
      1996 Annual Research Report
  • [Publications] T.Kohzuma: "Spectrosoopic and Electrochemical Studies on Active-site Transitions of the Type 1 Copper Protein Pseudoazurin from Achromobacter cycloclasres." J.Biol.Chem.270. 25733-25738 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] N.Shibata: "Orderly disposition of heterogeneous small subunits in D-ribulose-1,5-bisphosphate carboxylase/oxygenase from spinach." J.Biol.Chem.271. 26449-26452 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] N.Shibata: "Cristallization and preliminary crystalliographic studies of ribulose 1,5-bisphosphat carboxylase/oxygenase from red alga Galdieria partita with high specificity factor." J.Biochem.120. 1064-1066 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] C.Li: "Crystallization and Preliminary X-ray Studies of Plastocyanin from Silene Expressed in E.coli 1." Acta Cryst.D53. 129-130 (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] A.Toumoto: "Preliminary X-ray studies on a new crystal form of PHO4-DNA complex." Acta Cryst.D53. 103-104 (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] 芦田玉一 他: "Cystal structure analysis of phospholipase A2 from Trimeresurus flavoviridis venom at 1.4A resolution" J.Biochem.117. 730-740 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 山根隆 他: "Crystal structure of a new alkaline serine protease (M-protease) from Bacillussp. KSM-K16" Acta Crystallogr.D51. 199-206 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 佐藤能雅 他: "First hetero-bimetallic multifunctional asymmewtric catalyst" J.Am.Chem.Soc.117. 6194-6198 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 田中信夫 他: "Crystal structures of mutated 3-isopropyl malate dehydrogenase from Thermus thermophilus HB8 and their relationship to the thermostability of the enzyme" J.Biochem. 117. 408-413 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 月原富武 他: "Structure of metal sites of oxidized bovine heart cytochrome coxidase at 2.8A" science. 269. 1069-1074 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 三木邦夫 他: "Crystal structure of NADH-cytochrome b5 reductase from pig liver at 2.4A resolution" Biochemistry. 34. 2763-2767 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 芦田玉一 他: "Crystal structure analysis of phospholipase A2 from habu snake at 1.5A rersolution" J.Biochem.(印刷中). (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] 芦田玉一 他: "Crystal structure of a new alkaline serine protease (M-protease) from Bacillus sp.KSM-K16" Acta Crystallographica. D51(印刷中). (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] 安岡則武 他: "Location of active sites of NiFe Hydrogenase determined by the anomalous diffraction method" Acta Crystallographica. D50. 781-785 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 安岡則武 他: "バイオ分子の画像表示-グラフィックスワークステーション" 蛋白質核酸酵素. 39(増刊号). 1859-1863 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 佐藤能雅 他: "Structure of the allosteric regulatory enzyme of purine biosynthesis" Scieuce. 264. 1427-1433 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 月原富武 他: "Crystal structure analysis of [2Fe-2S]ferredoxin at 1.8A resolution" Acta Crystallographica. D50. 167-174 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 鈴木淳巨: "Crystallographic refinement of Bowman-Birk type protease inhibitor A-II from peanutat 2.3 A resolution" J.Mol.Biol.234. 722-734 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 鈴木淳巨: "Crystallization and preliminary X-ray studies on the trypsin inhibitor I-2 from wheat germ and its complex with trypsin" Acta Crystallogr.D49. 594-596 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Yasuoka 他: "On the active sites of hydrogenase from Desulfovibrio vulgaris Miyazaki F" Jpn.J.Appl.Phys.32. 553-555 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Tanaka 他: "Structure analysis of the myoglobin reconstituted with iron porphine" J.Biochem.268. 8935-8942 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 月原冨武: "超分子複合体のX線結晶構造解析" 蛋白質核酸酵素特集号、超分子システム研究法. 1247-1253 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Miki,Kunio 他: "Crystallization and preliminary X-ray diffraction studies of photolyase from cyanobacterium Anacystis nidulans" J.Mol.Biol.233. 167-169 (1993)

    • Related Report
      1993 Annual Research Report

URL: 

Published: 1993-04-01   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi