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X-ray Crystallographic Analysis of Bacteriophage Precursor Structures

Research Project

Project/Area Number 10044070
Research Category

Grant-in-Aid for Scientific Research (B).

Allocation TypeSingle-year Grants
Section一般
Research Field Biophysics
Research InstitutionTokyo Institute of Technology

Principal Investigator

ARISAKA Fumio  Fac. Biosci. & Bioeng. Tokyo Institute of Technology, Assoc. Professor, 生命理工学部, 助教授 (80133768)

Co-Investigator(Kenkyū-buntansha) ROSSMANN Mic  Dept. of Biological Sciences, Purdue大学, 教授
Project Period (FY) 1998 – 1999
Project Status Completed (Fiscal Year 1999)
Budget Amount *help
¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1999: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1998: ¥1,500,000 (Direct Cost: ¥1,500,000)
Keywordsmolecular chaperone / association equilibrium / sedimentation equilibrium / thermal denaturation / Gdn-HCl denaturation / infection / tail lysozyme / X-ray crystallography / バクテリオファージ / 尾部基盤 / 分子集合 / 分子認識 / 感染初期過程 / 構造変化 / リゾチーム
Research Abstract

As a step toward determination of the structure of the baseplate of bacteriophage T4 in atomic resolution, three precursor structures were chosen for crystallization, One of them was the initiation complex of the wedge of the baseplate, gp10-gp11 complex. It is considered to constitute the tail pins at the vertices of the hexagonal baseplate, which protrude from the bottom of the baseplate. Based on SDS-PAGE, Edman degradation of the complex and analytical ultracentrifugation, it was shown that the-complex is a hetero-hexamer with the subunit composition of (gp10)ィイD23ィエD2 (gp11)ィイD23ィエD2. Crystal of the complex suitable for X-ray analysis has not been obtained although small crystalls have been found. On the other hand, the second target for X-ray crystal analysis, i.e., gp5-gp27 complex, has been crystallized. Based on the results of sedimentation equilibrium and SDS-PAGE, the subunit structure of the complex was concluded to be hetero-nanomer, (gp5)ィイD23ィエD2(gp27)ィイD26ィエD2. The crystal yielded the resolution of approximately 2.5 A. At present, efforts are made to obatain isomorphous crystal containing selenomethionine. Gp5 is a structurally essential component, yet possesses lysozyme domain which locally digests peptidoglycan of the host E. coli upon infection so that the tail tube can reach the inner membrane. Concerning the third target protein, crystal has not been obtained, but we have recently succeeded in expressing a mutant protein which lacks the N-terminal seven residues that contain five negative charges. The mutant protein was shown to retain the native secondary and oligomeric structure. We plan to examine the possibility of crystallization of the mutant protein.

Report

(3 results)
  • 1999 Annual Research Report   Final Research Report Summary
  • 1998 Annual Research Report
  • Research Products

    (26 results)

All Other

All Publications (26 results)

  • [Publications] Masato USHIYAMA: "A 12-Porphyrin Construction of a System : Synthesis of Peptide Porphyrin with Multiple Histidines and the Aggregation Behavior in the Presence of Hemin"Bull. Chem. Soc. Japn. 72. 1351-1364 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kei-ichiro INAMORI: "A Newly Identififed Horseshoe Crab Lectin with Specificity for Blood Group A Antigen Recognizes Specific O-Antigens of Bacterial Lipopolysaccharides"J. Biol. Chem.. 274. 3272-3278 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Shuji KANAMARU: "C-terminal Fragment of the Precursor Tail Lysozyme of Bacteriophage T4 Stays as a Structural Component of the Baseplate after Cleavage"J. Bacteriology. 181. 2739-2744 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Mainul HOQUE: "Nuclear Transport of the major capsid protein is essential for Adeno-Associated virus Capsid Formation"J. Virol.. 73-9. 7912-7915 (19990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Shigeki TAKEDA: "Comparative Imaging of the T4 Phase Baseplate with Atomic Force and Electron Microscopy"Res. Comm. Biochem. & Mol. Biol.. (in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Li ZHAO: "Socichiometry and Inter-subunit Interaction of the Wedge Initiation Complex, Gp10-gp11 of Bacteriophage T4"Biochim. Biophys. Acta. (in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Li ZHAO: "Stoichimetry and Inter-subunit Interaction of the Wedge Initiation Complex, Gp 10-gp11 Bactriophage T4"Biochim. Biophys. Acta. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Shigeki TAKEDA: "Comparative Imaging of the T4 Phage Baseplate with Atomic Force and Electron Microscopy"Res. Comm. Biochem. & Mol. Biol.. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Masato USHIYAMA et al.: "A 12-Porphyrin Construction of a System : Synthesis of Peptide Porphyrin with Multiple Histidines and the Aggregation Behavior in the Presence of Hemin"Bull. Chem. Soc. Japn. 72. 1351-1364 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kei-ichiro INAMORI: "A Newly Identified Horseshoe Crab Lectin with Specificity for Blood Group A Antigen Recognizes Specific O-Antigens of Bacterial Lipopolysaccharides"J. Biol. Chem.. 274. 3272-3278 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Shunji KANAMARU: "C-terminal Fragment of the Precursor Tail Lysozyme of Bacteriophage T4 Stays as a Structural Component of the Baseplate after Cleavage"J. Bacteriology. 181. 2739-2744 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Mainul HOQUE: "Nuclear Transport of the major capsid protein is essential for Adeno-Associated Virus Capsid Formation"J. Virol.. 73-9. 7912-7915 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Shigeki TAKEDA: "Mapping of Functional Sites on the Primary Structure of the Tail Lysozyme of Bcteriophage T4 by Mutaion Analysis"Biochim. Biophys. Acta. 1384. 243-252 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Shigeki TAKEDA: "Discovery of the tail tube gene of bacteriophage Mu and sequence analysis of the sheath and tube genes"Biochim. Biophys. Acta. 1399. 88-92 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Masato USHIYAMA: "A 12-Porphyrin Construction of a System: Synthesis of Peptide Porphyrin with Multiple Histidines and the Aggregation Behavior in the Presence of Hemin"Bull. Chem. Soc. Japn. 72. 1351-1364 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Kei-ichiro INAMORI: "A Newly Identified Horseshoe Crab Leetin with Specificity for Blood Group A Antigen Recognizes Specific O-Antiges of Bacterial Lipopolysaccharides"J. Biol. Chem.. 274. 3272-3278 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Shuji KANAMARU: "C-terminal Fragment of the Precursor Tail Lysozyme of Bacteriophage T4 Stays as a Strucutral Component of the Baseplate after Cleavage"J. Bacteriology. 181. 2739-2744 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Mainul HOQUE: "Nuclear Transport of the major capsid protein is essential for Adeno-Associated Virus Capsid Formation"J. Virol.. 73-9. 7912-7915 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Shigeki TAKEDA: "Comparative Imaging of the T4 Phage Baseplate with Atomic Force and Electron Microscopy"Res. Comm. Biochem. & Mol. Biol.. (in press).

    • Related Report
      1999 Annual Research Report
  • [Publications] Li ZHAO: "Stoichiometry and Inter-subunit Interaction of the Wedge Initiation Complex, Gp10-gp11 of Bacteriophage T4"Biochim. Biophys. Acta. (in press).

    • Related Report
      1999 Annual Research Report
  • [Publications] Shigeki TAKEDA: "Mapping of Functional Sites on the Primary Structure of the Tail Lysozyme of Bacteriophage T4 by Mutation Analysis" Biochem.Biophys.Acta. 1384. 243-252 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Shigeki TAKEDA: "Discovery of the tail tube gene of bacteriophage Mu and sequence analysis of the sheath and tube genes" Biochim.Biophys.Acta. 1399. 88-92 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] 有坂文雄: "超遠心分析の基礎と新たな展開(1)" 蛋白質核酸酵素. 43(13). 2024-2032 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] 有坂文雄: "超遠心分析の基礎と新たな展開(2)" 蛋白質核酸酵素. 43(14). 2145-2152 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] 有坂文雄: "超遠心分析の基礎と新たな展開(3)" 蛋白質核酸酵素. 43(15). 2238-2244 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] 有坂文雄: "生体物質相互作用のリアルタイム解析実験法" シュプリンガーフェーラーク東京社, 268 (1998)

    • Related Report
      1998 Annual Research Report

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Published: 1998-04-01   Modified: 2016-04-21  

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