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Development of the ^<31>P-NMR Probe with ultra high sensitivity and analysis of phosphate binding proteins, related to signal transduction

Research Project

Project/Area Number 13558080
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Structural biochemistry
Research InstitutionTHE UNIVERSITY OF TOKYO

Principal Investigator

TANOKURA Masaru  University of Tokyo, Graduate School of Agricultural and sciences, Professor, 大学院・農学生命科学研究科, 教授 (60136786)

Co-Investigator(Kenkyū-buntansha) HARUYAMA Hideki  Sankyo Co., Ltd., Biomedical Research Laboratories, Vice-chief of laboratory (Reseacher), バイオメディカル研究所, 所次長(研究職)
Project Period (FY) 2001 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥14,100,000 (Direct Cost: ¥14,100,000)
Fiscal Year 2002: ¥5,000,000 (Direct Cost: ¥5,000,000)
Fiscal Year 2001: ¥9,100,000 (Direct Cost: ¥9,100,000)
Keywords^<31>P NMR / Probe / Phosphate binding protein / NMR / In vivo NMR / プローブ開発 / 生きた細胞 / 核磁気共鳴 / in vivo NMR / リンNMR / シグナル云達
Research Abstract

Although the sensitivity of ^<31>P-NMR (nuclear magnetic resonance) is a comparatively high and phosphate is existed mostly in forms, such as ATP, in the living body, ^<31>P is difficult for obtaining useful data because T_2 and T_1 are very short and long, respectively. Therefore, the sensitivity of the NMR measurement obtained so far has been a serious obstacle when advancing a research for performing an experiment to detect the signals from a sample containing single cells. However, the strength of the magnetic field used for NMR measurement goes up and probe technology was also improved in recent years, a possibility of measuring and analyzing by ^<31>P-NMR the single cells has also increased.
We have developed the probe equipment and the cell for detecting can carry out a direct monitoring the single cells in a condition making physiological environment. This probe can be improving a pulse for measurement and a receiver's dynamic range so that it can respond to short T_2 especially. Thus equipment hold the cell and the tissue in the state where it lived for a long time, it made it possible to measure the ^<31>P-NMR spectra. Moreover, it analyzed also about the milk, which is the mixture of the living substances used as the candidate for detection in this probe. It succeeded in assigning the phosphate compound as an ingredient in milk by applying a 2D ^1H-^<31>P NMR spectra in addition to ^1D ^<31>P NMR measurement, without pretreating.

Report

(3 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • Research Products

    (25 results)

All Other

All Publications (25 results)

  • [Publications] Sakai, N.et al.: "The three-dimensional structure of septum site-determining protein MinD from Pyrococcus horikoshii OT3 in complex with Mg-ADP."Structure. 9. 817-826 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yumoto, F.et al.: "Coordination structures of Ca^<2+> and Mg^<2+> Akazara scallop troponin C in solution : FTIR spectroscopy of side-chain COO^- groups."Fur.J.Biochem.. 268. 6284-6290 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Kato, Y.et al.: "Determinants of ligand specificity in groups I and IV WW domains as studied by surface plasmon resonance and model building."Fur.J.Biochem.. 277. 10173-10177 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Sawano, Y.et al.: "Characterization of genomic sequence coding for bromelain inhibitors in pineapple and expression of its recombinant isoform."J.Biol.Chem.. 277. 28222-28227 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Iwasaki, W.et al.: "Metal-free and Ca^<2+>-bound structures of a multidomain EF-hand protein, CBP40, from the lower eukaryote, Physarum polycephalum."Structure. 11. 75-85 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hatano, K.et al.: "Nuclear magnetic resonance studies on the pK_a values and interactions of ionizable groups in bromelain inhibitor VI from pineapple stem."Biol.Chem.. 384. 93-104 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 田之倉 優: "生化学&ポストゲノム最前線.タンパク質の機能解析の実現に向けて"日経サイエンス. 1 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Sakai, N., Yao, M., Itou, H., Watanabe, N., Yumoto, F., Tanokura, M., Tanaka, I.: "The three-dimensional stricture of septum site-determining protein MinD from Pyrococcus horikoshii OT3 in complex with Mg-ADP."Structure. 9. 817-826 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yumoto, F., Nara, M., Kagi, H., Iwasaki, W., Ojima, T., Nishita, K., Nagata, K., Tanokura, M.: "Coordination strictures of Ca^<2+>+ and Mg^<2+>+ in Akazara scallop troponin C in solution : FTIR spectroscopy of side-chain COO^-groups."Eur.J.Biochem.. 268. 6284-6290 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Kato, Y., Ito, M., Kawai, K., Nagata, K, Tanokura, M.: "Determinants of ligand specificity in groups I and IV WW domains as studied by surface plasmon resonance and model building."J.Biol.Chem.. 277. 10173-10177 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Sawano, Y., Muramatsu, T., Hatano, K., Nagata, K., Tanokura, M.: "Characterization of genomic sequence coding for bromelain inhibitors in pineapple and expression of its recombinant isoform."J.Biol.Chem.. 277. 28222-28227 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Iwasaki, W., Sasaki, H., Nakamura, A., Kohama K., Tanokura, M.: "Metal-free and Ca^<2+>-bound strictures of a multidomain EF-hand protein, CBP40, from the lower eukaryote, Physarum polycephalum."Structure. 11. 75-85 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hatano, K., Kojima, M., Tanokura, M., Takahashi, K.: "Nuclear magnetic resonance studies on the pK_a values and interactions of ionizable groups in bromelain inhibitor VI from pineapple stem."Biol.Chem.. 384. 93-104 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Kato, Y., Ito, M., Kawai, K., Nagata, K., Tanokura, M: "Determinants of ligand specificity in groups I and IV WW domains as studied by surface plasmon resonance and model building"Journal of Biological Chemistry. 277・12. 10173-10177 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Sawano, Y., Muramatsu, T., Hatano, K., Nagata, K., Tanokura, M: "Characterization of genomic sequence coding for bromelain inhibitors in pineapple and expression of its recombinant isoform"Journal of Biological Chemistry. 277・31. 28222-28227 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Iwasaki, W., Sasaki, H., Nakamura, A., Kohama K., Tanokura, M.: "Metal-free and Ca^<2+> -bound structures of a multidomain EF-hand protein, CBP4O, from the lower eukaryote, Physarum polycephalum"Structure. 11・1. 75-85 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Hatano, K., Kojima, M., Tanokura, M., Takahashi, K: "Nuclear magnetic resonance studies on the pK_a values and interactions of ionizable groups in broinelain inhibitor VI from pineapple stem"Biological Chemistry. 384・1. 93-104 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Maeda, M., Takeuchi, K., Kojima, M., Tanokura, M., Kimura, K., Amemiya, Y., Kihara, H., Takahashi, K: "Kinetic studies of unfolding process of aspergillopepsinn II by pH-jump methods"Biochemical and Biophysical Research communications. 301・3. 745-750 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Katayama, H., Nagata, K., Ohira, T., YF., Tanokura, M., Nagasawa, H.: "The solution structure of molt-inhibiting hormone from the kuruma prawn Marsupenaeus japonicus"Journal of Biological Chemistry. (in press). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] 田之倉優, 阿久津秀雄, 村松知成: "生化学キーノート(Hammes, B.D, Hooper, N.M.著)"シュプリンガーフェアラーク東京. 464 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] 田之倉優, 村松知成, 八木澤 仁: "分子生物学キーノート(Turner, P.C., McLennan, A.G., Bates, A.D., White, M.R.H.著)"シュプリンガーフェアラーク東京. 398 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Sakai, N., Yao, M., Itou, H., Watanabe, N., Yumoto, F., Tanokura, M., Tanaka, I.: "The three-dimensional structure of septum site-determiining protein MinD from Pyrococcus horikioshii OT3 in complex with Mg-ADP"Structure. 9・9. 817-826 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Yumoto, F., Nara, M., Kagj, H., Iwasaki, W., Ojima, T., Nishita, K., Nagata, K., Tanokura, M.: "Coordination structures of Ca^<2+> and Mg^<2+> in Akazara scallop troponin C in solution : FTIR sectroscopy of side-chain COO Grous"European Jounal of Biochemistry. 268. 6284-6290 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Tanaka, S., Ataka, M., Kubota, T., Soga, T., Homma K., Lee W.C., Tanokura M.: "Thee effect of amphiphillic additives on the growth and morphology of Aspergillus niger acid proteinase A crystals"Journal of Crystal Growth. 234. 247-254 (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] Kato, Y., Ito, M., Kawai, K., Nagata, K., Tanokura, M.: "Determinants of ligand specificity in groups I and IV WW domains as studied by surface plasmon resonance and model building"Journal of Biological Chemistry. 277(in press). (2002)

    • Related Report
      2001 Annual Research Report

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Published: 2001-04-01   Modified: 2016-04-21  

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