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Development of high-throughput mass-spec based sequencing and its application to global analysis of protein phosphorylation

Research Project

Project/Area Number 11558082
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Structural biochemistry
Research InstitutionRIKEN (2000-2001)
Fujita Health University (1999)

Principal Investigator

TANIGUCHI Hisaaki  RIKEN, Postranlational Modifications and Dynamic Regulation Research Team, Team Leader, 翻訳後修飾による動的調節機構研究チーム, チームリーダー(研究職) (10257636)

Co-Investigator(Kenkyū-buntansha) MATSUBARA Mamoru  RIKEN, Postranlational Modifications and Dynamic Regulation Research Team, Researcher, 翻訳後修飾による動的調節機構研究チーム, 連携研究員 (90288481)
千谷 晃一  藤田保健衛生大学, 総合医科学研究所, 教授 (60179942)
Project Period (FY) 1999 – 2001
Project Status Completed (Fiscal Year 2001)
Budget Amount *help
¥12,000,000 (Direct Cost: ¥12,000,000)
Fiscal Year 2001: ¥2,800,000 (Direct Cost: ¥2,800,000)
Fiscal Year 2000: ¥2,800,000 (Direct Cost: ¥2,800,000)
Fiscal Year 1999: ¥6,400,000 (Direct Cost: ¥6,400,000)
Keywordsamino acid sequencing / proteome / mass analysis / mass spectrometry / protein / peptide / マウスペクトロメトリー / プロテオーム解析 / ゲノム解析 / 翻訳後修飾 / 蛋白質リン酸化 / シグナル伝達
Research Abstract

The aims of the research are :
1) To establish, mass mapping method to identify proteins in the genome database with the information obtained by mass spectrometry of peptides generated from 2D gel spots.
2) TO establish a modification-specific method to detect phosphorylated or acetylated peptides.
3) To apply thus established methods to the global analysis of signaling complexes to elucidate the functions of "unknown" proteins.
For these porpoises, we have established an analysis system, in which gel-spotting and excision, mass measurements and database search are performed automatically. To achieve a high-throughput analysis, the sample-pretreatment robot was purchased by the fund, and modified to make automatic desalting and targeting on the sample plate possible. We have also established an analysis system with which specific detection of phosphopeptides are possible by parent scan in the triple-quad mass spectrometer. The high-throughput identification of more than 30 proteins in a single sample became possible by LC/MS analysis. In this way, we could achieve the high-throughput mass sequencing.

Report

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

    (28 results)

All Other

All Publications (28 results)

  • [Publications] Takasaki, A.: "Identification of the calmodulin-binding domain of neuron-specific protein kinase C substrate protein CAP-22/NAP-22"J. Biol. Chem.. 274. 11848-11853 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Manenti, S.: "Phosphorylation of the myristoylated PKC substrate MARCKS by the cyclin E-Cdk2 complex in vitro"Biochem. J.. 340(Pt3). 775-782 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Taniguchi, H.: "Protein myristoylation in protein-lipid and protein-protein interactions (Review)"Biophys. Chem.. 82. 129-137 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Morimatsu, M.: "The molecular structure of the fastest myosin from green algae, Chara"Biochem Biophys Res Commun.. 270. 147-152 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Sano, K.I.: "Amino-acid replacements in an internal region of tropomyosin alter the properties of the entire molecule"Eur. J. Biochem.. 267. 4870-4877 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Konishi, K.: "Tyrosine phosphorylation sites of protein kinase C δ in H_2O_2-treated cells and its implication in the regulation of the kinase activity"Proc. Natl. Acad. Sci.. 98. 6587-6592 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] 伊藤隆司, 谷口寿章: "ポストシークエンスのゲノム科学第3巻プロテオミクス"中山書店. 238 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Takasaki, A., et al: "Identification of the calmodulm-binding domain of beuron-specific proteinkinase C substrate protein CAP-22"J. Biol. chem.. 274. 11848-11853 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Manenti, S., et al: "Phosphorylation of the myristoylated PKC substare MARCKS by the cyclin E-Cdk2 complex in vitro"Biochem. J.. 340(Pt3). 775-782 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Taniguchi, H: "Protein myristoylation in protein-lipid and protein-protein interactions(Review)"Biophys. Chem.. 82. 129-137 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Morimatsu, M. et al: "The molecular structure of the fastest myosin from green algae, Chara"Biodhem Biophys Res Commun.. 267. 147-152 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Sano, K., et al: "Amino-acid replacements in an internal region of tomyosin alter the properties of the entire molecule"Eur. J. Biochem.. 67. 4870-4877 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Konishi, K., et al: "Tyrosine phosphorylation sites of protein kinase.C in H2O2-treated cells and its implication in the regulation of the kinase activity."Proc. Natl. Acad. Sci., USA. 98. 6587-6592 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Konishi, K: "Tyrosine phosphorylation sites of protein kinase C in H_2O_2-treated cells and its implication in the regulation of the kinase activity"Proc. Natl. Acad. Sci., USA. 98. 6587-6592 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 谷口寿章: "プロテオミクスの現状と展開"蛋白質核酸酵素. 46. 2603-2607 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Morimatsu.M., et al.: "The molecular structure of the fastest myosin from green algae, Chara."Biochem Biophys Res Commun. 270. 147-152 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Sano,K.I., et al.: "Amino-acid replacements in an internal region of tropomyosin alter the properties of the entire molecule."Eur.J.Biochem.. 267. 4870-4877 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 谷口寿章: "シナプス形成と伝達物質放出におけるプロリン指向性キナーゼの役割"蛋白質核酸酵素増刊「神経回路と機能発達」. 45. 436-441 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 谷口寿章: "四重極マスフィルター型質量分析計とその応用"蛋白質核酸酵素. 45. 1865-1871 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 谷口寿章: "プロテオーム解析の現状と展望"医学のあゆみ. 195. 927-930 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 伊藤隆司,谷口寿章: "ポストシークエンスのゲノム科学第3巻プロテオミクス"中山書店. 238 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Uedaira, H., et al.: "Domain organization of flagellar hook protein from Salmonella typhimurium"FEBS Lett.B. 421. 203-207 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Takasaki, A., et al.: "Identification of the calmodulin-binding domain of neuron-specific protein kinase C substrate protein CAP-23/NAP22 : Direct involvement of protein myristoylation in calmodulin-target protein interaction"J. Biol. Chem.. 274. 11848-11853 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Manenti, S., et al.: "Phosphorylation of the myristoylated PKC substrate MARCKS by the cyclin E-Cdk2 complex in vitro"Biochem. J.. 340. 775-782 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Taniguchi, H.: "Protein myristoylation in protein-lipid and protein-protein interactions (Review)"Biophys. Chem.. 82. 129-137 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] 谷口寿章: "質量分析法のたんぱく質構造解析への応用とプロテオーム解析"実験医学. 17. 2550-2554 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Yamauchi, E. and Taniguchi, H.: "Neural development (Uemura, K. Kawamura, K. and Yazaki, T., eds.)"Springer-Verlag, Tokyo. 416-420 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Yamauchi, E., et al.: "Neural development (Uemura, K. Kawamura, K. and Yazaki, T., eds.)"Springer-Verlag, Tokyo. 421-424 (1999)

    • Related Report
      1999 Annual Research Report

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Published: 1999-04-01   Modified: 2016-04-21  

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