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Development of New Methodologies for Protein-Interaction Analysis

Research Project

Project/Area Number 10179101
Research Category

Grant-in-Aid for Scientific Research on Priority Areas (A)

Allocation TypeSingle-year Grants
Review Section Biological Sciences
Research InstitutionOsaka University (2000-2001)
Yokohama National University (1998-1999)

Principal Investigator

AKUTSU Hideo  Institute for Protein Research, Osaka University, Professor, 蛋白質研究所, 教授 (60029965)

Co-Investigator(Kenkyū-buntansha) KATAOKA Mikio  Nara Institute of Technology, Professor, 教授 (30150254)
SATO Mamoru  Yokohama City University, Professor, 大学院・総合理学研究科, 教授 (60170784)
KAINOSHO Masatsune  Tokyo Metropolitan University, Professor, 大学院・理学研究科, 教授 (20137029)
YAMAZAKI Toshio  Institute for Protein Research, Osaka University, Associate Professor, 蛋白質研究所, 助教授 (60273710)
TSUKIHARA Tomitake  Institute for Protein Research, Osaka University, Professor, 蛋白質研究所, 教授 (00032277)
相本 三郎  大阪大学, 蛋白質研究所, 教授 (80029967)
Project Period (FY) 1998 – 2001
Project Status Completed (Fiscal Year 2001)
Budget Amount *help
¥132,200,000 (Direct Cost: ¥132,200,000)
Fiscal Year 2001: ¥30,800,000 (Direct Cost: ¥30,800,000)
Fiscal Year 2000: ¥34,700,000 (Direct Cost: ¥34,700,000)
Fiscal Year 1999: ¥32,000,000 (Direct Cost: ¥32,000,000)
Fiscal Year 1998: ¥34,700,000 (Direct Cost: ¥34,700,000)
KeywordsProtein interaction / Protein segmental labeling / NMR / Solution structure determination / Magic angle sample spinning / Solid state NMR / Ab initio X-ray crystallography / Thioester method / 部分セグメント標識法 / 立体・位置特異的重水素化 / 固体高分解能多次元NMR / 安定同位体標識 / X線溶液散乱法 / 電子顕微鏡単粒子解析 / 第3世代放射光 / 多波長異常分散法 / タンパク質相互作用 / 情報伝達 / X線結晶構造解析 / 電子顕微鏡 / 同位体標識 / X線小角散乱
Research Abstract

1) A new segmental isotope labeling method has been developed for structural analysis of large proteins. This method was successfully applied to RNA polymerase α subunit C-domain, maltose-binding protein, ATPase β-subunit.
2) A series of new techniques in solid state NMR have been developed for structural analysis of large proteins and membrane proteins that are unsuitable for the analysis by X-ray crystallography and solution NMR.
3) An Ab initio method for X-ray crystal analysis have been developed for analysis of huge biological systems.
4) A thioester method has been developed for chemical synthesis of proteins to investigate protein-protein interactions.

Report

(5 results)
  • 2001 Annual Research Report   Final Research Report Summary
  • 2000 Annual Research Report
  • 1999 Annual Research Report
  • 1998 Annual Research Report
  • Research Products

    (38 results)

All Other

All Publications (38 results)

  • [Publications] TozaWa, K.: "Functions and ATP-binding responses of the twelve histidine residues in the TF_1-AT Pase β subunit"J.Biochem.. 130. 527-533 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Hanaoka, S.: "NMR Structure of the hRap1 Myb Motif Reveals a Canonical Three-hekix Bundle Lacking the Positive Surface Cnarge Typical of Myb DNA-Binding Domains"J.Mol.Biol.. 312. 167-175 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Riek, R.: "[^<13>C,^<13>C]-and [^<13>C,^1H]-TROSY in a Triple resonance Experiment for Ribose-base Intrabase Correlations in Nuclei Acids"J.Am.Chem.Soc.. 123. 658-664 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Yuzawa, S.: "Solution structure of Grbz reveals extensive flexibility necessary of target recognition"J.Mol.Biol.. 306. 527-537 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Ida, K.: "The 1.55 A resolution structure of Nicotiana alata S_<F11>-RNase associated with gametophytic self-incompatibility"J.Mol.Biol.. 314. 103-112 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Tanaka, H.: "X-ray Crystalline Structure of Pyrnolidine Carboxyl peptidase from a Hyperthenmophile, Pyrococcus furiosus, and Its Cys-Free Mutant"J.Biochem.. 130. 107-118 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Tozawa, K.: "Functions and ATP-binding responses of the twelve histidine residues in the TF_1-AT Pase β subunit"J. Biochem.. Vol. 130. 527-533 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Hanaoka, S.: "NMR Structure of the hRap1 Myb Motif Reveals a Canonical Three-hekix Bundle Lacking the Positive Surface Cnarge Typical of Myb NDA-Binding Domains"J. Mol. Biol.. Vol. 312. 167-175 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Riek, R.: "[^<13>C, ^<13>C]-and [^<13>C, ^1H]-TROSY in a Triple resonance Experiment for Ribose-base intrabase Correlations in Nuclei Acids"J. Am. Chem. Soc.. Vol. 123. 658-664 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Yuzawa, S.: "Solution structure of Grbz reveals extensive flexibility necessary of target recognition"J. Mol. Biol.. Vol. 306. 527-537 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Ida, K.: "The 1.55 A resolution structure of Nicotiana alata S_<F11>-Rnase associated with gametophytic self-incompatibility"J. Mol. Biol.. Vol. 314. 103-112 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Tanaka, H.: "X-ray Crystalline Structure of Pymolidine Carboxyl peptidase from a Hyperthenmophile, Pyrococcus furiosus, and Its Cys-Free Mutant"J. Biochem.. Vol. 130. 107-118 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] K.Tozawa 外5名: "Functions and ATP-binding responses of the twelve histidine residues in the TF_1-ATPase β subunit"J. Biochem.. 130. 527-533 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] S.Hanaoka 外6名: "NMR Structure of the hRap1 Myb Motif Reveals a Canonical Three-helix Bundle Lacking the Positive Surface Change Typical of Myb DNA-Binding Domains"J. Mol. Biol.. 312. 167-175 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] R.Riek 外4名: "[^<13>C,^<13>C] -and [^<13>C,^1H] -TROSY in a Triple Resonance Experiment for Ribose -base Intrabase Correlations in Nuclei Acids"J. Am. Chem. Soc.. 123. 658-664 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] S.Yuzawa 外8名: "Solution structure of Grb2 reveals extensive flexibility necessary for target recognition"J. Mol. Biol.. 306. 527-537 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] K.Ida 外8名: "The 1,55 A resolution structure of Nicotiana alata SF11-RNase associated with gametophytic self-incompatibility"J. Mol. Biol.. 314. 103-112 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] H.Tanaka 外6名: "X-ray Crystalline Structure of Pyrrolidine carboxyl peptidase from a Hyperthermophile, Pynococcus furiosus, and Its Cys-Free Mutant"J. Biochem.. 130. 107-118 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] T.Fujiwara 外2名: "Compound radio-frequency driven recoupling pulce sequences for efficient vaguetization transfer by nomounaclear dipolar interaction under magic angle spinning coudition"J.Magu.Reson.. 145. 73-83 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] M.Mishima 外2名: "Intermolacular ^<31>P-^<15>N and ^<31>P-^1H scalar coupling across hydrogen bouds formed between a protein and a uncleotide"J.Am.Chem.Soc.. 122. 5883-8884 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] N.Utsunominga-Tate 外6名: "Reelin Molecules Assemble Together to Foum a large Protein Complex, which is Inhibited by the Functional-blocking CR-50 Antibody"Pnoc.Natl.Acad.Sci.USA. 97. 9729-9734 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] T.Oka 外5名: "Time-resolved X-ray diffraction reveale multiple conformations in the M-N transition of the bacteriorhodopciu photocycle"Proc.Natl.Acad.Sci.USA. 97. 14278-14282 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] S.Mitsuhashi 外8名: "X-ray structure of b-carbonic anhydrase from the red alga, porphyridium purpureum., reveals a novel catalytic site for CO_2 hydration"J.Biol.Chem.. 275. 5521-5526 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] T.Otomo 外4名: "Structure of the heterodimenic complex between CAD domains of CAD and ICAD"Nature Strut.Biol.. 7. 658-662 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] K. Nomura: "Determination of the Complete Structure of a Uniformly Labeled Molecule by Rotational Resonance Solid-State NMR in the Tilted Rotational Frame"J. Am. Chem. Soc.. 121. 4064-4065 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] C. Fernandz: "Conformational Changes of the BS2 Operator DNA Complex Formation with the Antennapedia Homeodomain Assessed by ^<13>C/^<15>N Labeling of the DNA"J. Mol. Biol.. 292. 609-617 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Nagata T.: "Structure and interactions with RNA of the N-terminal UUAG-specific domain of hnRNP D0"J. Mol. Biol.. 287. 221-237 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Nagata T.: "Structure, backbone dynamics and interactions with RNA of the C-terminal RNA-binding domain of a mouse neural RNA-binding protein, Musashi1"J. Mol. Biol.. 287. 315-330 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Takanori Otomo: "NMR observation of selected segments in a larger proteins : central-segment isotope labeling through intein-mediated ligation"Biochemistry. 38. 16040-16044 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] H. Yagi: "Functional conformational changes in the TF, ATPase βsubunit probed by twelve tyrosine residues"Biophys. J.. 77. 2175-2183 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] 阿久津秀雄・河野敬一: "多核NMR 自然を複眼で見る"廣川書店. 147 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] T.Fujiwara: "Multidimensional solid-state NMR for determining dihedral angle from the correlation of ^<13>C-^1H and ^<13>C-^<13>C dipolar interactions under magic-angle spinning conditions." J.Chem.Phys.109. 2380-2309 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] K.Pervushin.: "NMR Scalar Coupling Across Watson-Crick Base Pair Hydrogen Bonds in DNA Observed by TROSY" Proc.Natl.Acad.Sci.USA. 95. 14147-14151 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] K.Haneda,: "Chemo-enzymatic synthesis of calcitonin derivatives containing N-linked origosaccharides" Bioorg.Medi.Chem.Lett.8. 1303-1306 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] S.Yoshikawa: "Redox-Coupled Crystal Structural Change in Bovine Herart Cytochrome c Oxidase." Science. 280. 1723-1729 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] K.Ihara: "Evolution of the Archaeal Rhodopsins: Evolution Rate Changes by Gene Duplication and Functional Differentiation." J.Mol.Biol.285. 163-174 (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Masayuki Oda: "Thermodynamics of specific and non-specific DNA binding by the c-Myb DNA-binding domain." J.Mol.Biol.276. 571-590 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] 片平正人: "核酸の構造と機能、生命薬学テキストシリーズ:物理化学上下(分担)" 共立出版(印刷中), (1999)

    • Related Report
      1998 Annual Research Report

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Published: 1998-04-01   Modified: 2018-03-28  

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