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Development of the reliable methods to detect the interaction of proteins and ligands using diffusion of molecules

Research Project

Project/Area Number 15550076
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Analytical chemistry
Research InstitutionDepartment of Chemistry, College of Science and Technology, Meisei University (2005)
Tokyo Metropolitan University (2003-2004)

Principal Investigator

TASHIRO Mitsuru  Meisei University, Chemistry, Associate Professor, 理工学部, 助教授 (40315750)

Co-Investigator(Kenkyū-buntansha) MACHINAMI Tomoya  Meisei University, Chemistry, Professor, 理工学部, 教授 (50247160)
ONO Akira  Kanagawa University, Applied Chemistry, Professor, 工学部, 教授 (10183253)
Project Period (FY) 2003 – 2005
Project Status Completed (Fiscal Year 2005)
Budget Amount *help
¥2,800,000 (Direct Cost: ¥2,800,000)
Fiscal Year 2005: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2004: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2003: ¥1,200,000 (Direct Cost: ¥1,200,000)
Keywordsdrug screening / nuclear magnetic resonance / molecular diffusion / intermolecular interaction / 結合水 / 核磁気共鳴法(NMR) / コールドスプレーイオン化質量分析法
Research Abstract

There is a growing need for efficient methods, capable of reliably identifying compounds in a relatively short time span with desired binding affinity. In designing NMR-based screening methods for lead compounds, it is critical to develop the technique capable of screening a large pool of new drug candidates and identifying lead compounds with high affinity towards the target proteins. an improved version of the WaterLOGSY sequence is proposed, and its efficiency of identifying a ligand bound to a protein is compared with that of STD and NOE-pumping techniques. The effectiveness of this NMR-based screening method is demonstrated using the RNase T_1-inhibitor system.
Water-LOGSY experiment was improved by incorporating the double pulsed field gradient spin-echo (DPFGSE) sequence. DPFGSE sequence provides superior selective excitation without phase distortion problems that are usually associated with conventional selective excitation pulses. In the WaterLOGSY spectra, the positive signals from the inhibitors bound to RNase T_1 were clearly observed for the H8 proton of 5'-GMP and the H2 and H8 protons of 3'-AMP. These signals were not observed in the other techniques, which indicate the effectiveness of the DPFGSE-WaterLOGSY.
Practical aspects of several ROESY pulse sequences have been investigated to identify bound water molecules of the non-labeled saccharide. To increase the selectivity of water resonance excitation, very weak rf strengths for the selective 180° pulses, corresponding to pulse widths of 500 ms, was used in DPFGSE. Several Cross peaks arising from bound water proton-proton of saccharide spin-spin cross relaxation are observed, and a binding water was identified by NMR.

Report

(4 results)
  • 2005 Annual Research Report   Final Research Report Summary
  • 2004 Annual Research Report
  • 2003 Annual Research Report
  • Research Products

    (28 results)

All 2006 2005 2004 Other

All Journal Article (23 results) Publications (5 results)

  • [Journal Article] Mercury^II -mediated fonnation of thymine-Hg^II -thymine base pairs in DNA deplexes2006

    • Author(s)
      Y.Miyake, H.Togashi, M.Tashiro, H.Yamaguchi, S.Oda, et al.
    • Journal Title

      J. Am. Chem. Soc. 128

      Pages: 2172-2173

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Spectroscopic characterization of 2-isopropylinalic acid-aluminum(III) complex2006

    • Author(s)
      M.Tashiro, T.Fujimoto, T.Suzuki, K.Furihata, T.Machinami, E.Yoshimura
    • Journal Title

      J. Inorg. Biochem. 100

      Pages: 201-205

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Mercury ^<II> -mediated formation of thymine-Hg ^<II> -thymine base pairs in DNA deplexes2006

    • Author(s)
      Y.Miyake, H.Togashi, M.Tashiro, H.Yamaguchi, S.Oda, M.Kudo, Y.Tanaka, Y.Kondo, R.Sawa, T.Fujimoto, T.Machinami, A.Ono
    • Journal Title

      J.Am.Chem.Soc. 128

      Pages: 2172-2173

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Spectroscopic characterization of 2-isopropylmalic acid-aluminum(III) complex2006

    • Author(s)
      M.Tashiro, T.Fujimoto, T.Suzuki, K.Furihata, T.Machinami, E.Yoshimura
    • Journal Title

      J.Inorg.Biochem. 100

      Pages: 201-205

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Mercury^<II> -mediated formation of thymine-Hg^<II> -thymine base pairs in DNA deplexes2006

    • Author(s)
      Y.Miyake, H.Togashi, M.Tashiro, H.Yamaguchi, S.Oda, et al.
    • Journal Title

      J.Am.Chem.Soc. 128

      Pages: 2172-2173

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Spectroscopic characterization of 2-isopropylmalic acid-aluminum (III) complex2006

    • Author(s)
      M.Tashiro, T.Fujimoto, T.Suzuki, K.Furihata, T.Machinami, E.Yoshimura
    • Journal Title

      J.Inorg.Biochem. 100

      Pages: 201-205

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Observation of 1,6-anhydro-β-D-maltose and 1,6-anhydro-β-D-glucopyranose bomplexed with rubidium by NMR spectroscopy and electrospray ionization mass spectrometry2005

    • Author(s)
      T.Fujimoto, S.Sakuraki, A.Tsutsui, K.Furihata, T.Machinami, M.Tashiro
    • Journal Title

      Anal. Sci. 21

      Pages: 1245-1247

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Practical Aspects of ROESY Experiments for Identification of Bound Waters in the Cyclic Tetrasaccharide2005

    • Author(s)
      K.Furihata, T.Fujimoto, A.Tsutsui, T.Machinami, M.Tashiro
    • Journal Title

      Magn. Reson、 Chem. 43

      Pages: 1044-1048

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Identification of bound water molecules in the cyclic tetrasaccharide cyclo-{'6}-a-D-Glop-(l' 3)-a-D-Glop-(l' 6)-a-D-Glop-(1' 3)-a-D-Glop-(l' )2005

    • Author(s)
      K.Furihata, T.Fujimoto, A.Tsutsui, T.Machinami, M.Tashiro
    • Journal Title

      Carholwdr. Res. 340

      Pages: 2060-2063

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Observation of Water Molecules Bound to a Protein Using Cold-Spray Ionization Mass Spectrometry2005

    • Author(s)
      Y.Sei, S.Shimotakahara, J.Ishii, H.Shindo, H.Seki, K.Yamaguchi, M, Tashiro
    • Journal Title

      Anal. Sci. 21

      Pages: 449-451

    • NAID

      10015513763

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Observation of 1,6-anhydro-β-D-maltose and 1,6-anhydro-β-D-glucopyranose complexed with rubidium by NMR spectroscopy and electrospray ionization mass spectrometry2005

    • Author(s)
      T.Fujimoto, S.Sakurai, A.Tsutsui, K.Furihata, T.Machinami, M.Tashiro
    • Journal Title

      Anal.Sci. 21

      Pages: 1245-1247

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Practical Aspects of ROESY Experiments for Identification of Bound Waters in the Cyclic Tetrasaccharide2005

    • Author(s)
      K.Furihata, T.Fujimoto, A.Tsutsui, T.Machinami, M.Tashiro
    • Journal Title

      Magn.Reson.Chem. 43

      Pages: 1044-1048

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Annual Research Report 2005 Final Research Report Summary
  • [Journal Article] Identification of bound water molecules in the cyclic tetrasaccharide cyclo-{→6}-α-D-Glcp-(1→3)-α-D-Glcp-(1→6)-α-D-Glcp-(1→3)-α-D-Glcp-(1→)2005

    • Author(s)
      K.Furihata, T.Fujimoto, A.Tsutsui, T.Machinami, M.Tashiro
    • Journal Title

      Carbohydr.Res. 340

      Pages: 2060-2063

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Observation of Water Molecules Bound to a Protein Using Cold-Spray Ionization Mass Spectrometry2005

    • Author(s)
      Y.Sei, S.Shimotakahara, J.Ishii, H.Shindo, H.Seki, K.Yamaguchi, M.Tashiro
    • Journal Title

      Anal.Sci. 21

      Pages: 449-451

    • NAID

      10015513763

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Application of NMR screening techniques for observing a ligand binding with a protein receptor2005

    • Author(s)
      S.Shimotakahara, K.Furihata, M.Tashiro
    • Journal Title

      Magn.Reson.Chem. 43

      Pages: 69-72

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary 2004 Annual Research Report
  • [Journal Article] Observation of 1,6-anhydro-β-D-maltose and 1,6-anhydro-β-D-glucopyranose complexed with rubidium by NMR spectroscopy and electrospray ionization mass spectrometry2005

    • Author(s)
      T.Fujimoto, S.Sakuraki, A.Tsutsui, K.Furihata, T.Machinami, M.Tashiro
    • Journal Title

      Anal.Sci. 21

      Pages: 1245-1247

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Identification of bound water molecules in the cyclic tetrasaccharide cyclo-{→6}-α-D-Glcρ-(l→3)-α-D-Glcρ-(l→6)-α-D-Glcρ-(l→3)-α-D-Glcρ (1→)2005

    • Author(s)
      K.Furihata, T.Fujimoto, A.Tsutsui, T.Machinami, M.Tashiro
    • Journal Title

      Carbohydr.Res. 340

      Pages: 2060-2063

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Observation of Water Molecules Bound to a Protein Using Cold-Spray Ionization Mass Spectrometry2005

    • Author(s)
      Y.Sei, S.Shimotakahara, J.Ishii, H.Shindo, H.Seki, K.Yamaguchi, M.Tashiro
    • Journal Title

      Anal.Sci 21

      Pages: 449-451

    • NAID

      10015513763

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Thermal Unfolding of Ribonuclease TI by Multi-Dimensional NMR Spectroscopy2005

    • Author(s)
      H.Matsuura, S.Shimotakahara, M.Tashiro, K.Takahashi et al.
    • Journal Title

      Biol.Chem. 385

      Pages: 1157-1164

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Application of the ^<19>F NMR Technique to Observe Binding of the General Anesthetic Halothane to Human Serum Albumin2004

    • Author(s)
      K.Shikii, S.Sakurai, H.Utsumi, H.Seki, M.Tashiro
    • Journal Title

      Anal Sci. 20

      Pages: 1475-1477

    • NAID

      10013657744

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Computational and NMR Analyses for the Identification of Bound Water Molecules in Ribonuclease T_12004

    • Author(s)
      S.Shimotakahara, S.Hojo, K.Furihata, M.Tashiro
    • Journal Title

      Anal.Sci. 20

      Pages: 1471-1474

    • NAID

      10013657713

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Application of difference NOE-pumping NMR technique and coldspray ionization mass spectrometry to identify a ligand binding with a protein receptor2004

    • Author(s)
      H.Seki, Y.Sei, S.Shimotakahara, H.Utsumi, K.Yamaguchi, M.Tashiro
    • Journal Title

      Anal.Sci. 20

      Pages: 1467-1470

    • NAID

      10013657686

    • Related Report
      2004 Annual Research Report
  • [Journal Article] コールドスプレーイオン化質量分析2004

    • Author(s)
      清 悦久, 関 宏子, 田代 充, 藤田 誠, 山口 健太郎ほか
    • Journal Title

      分析化学 53

      Pages: 457-474

    • NAID

      110002905389

    • Related Report
      2004 Annual Research Report
  • [Publications] K.Furihata, S.Shimotakahara, M.Tashiro: "Improvement of double pulsed field gradient spin-echo ROESY experiments for identification of bound waters"Magnetic Resonance in Chemistry. 41. 955-958 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] H.Utsumi, H.Seki, K.Yamaguchi, M.Tashiro: "Segment identification of a ligand binding with a protein receptor using multidim ensional T1ρ-, diffusion-filtered and diffusion-ordered NOESY experiments"Analytical Science. 19. 1441-1443 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] M.Tashiro, N.Ishida, S.Shimotakahara, S.Tanabe, A.Okubo: "Ontogenetic changes of the water status in the heated Quail's egg as studied by nuclear magnetic resonance imaging"Analytical Science. 19. 933-936 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] M.Tashiro, et al.: "NMR structure of ubiquitin-like domain in PARKIN : gene product of familial Parkinson's disease"J.Biomol.NMR. 25. 153-156 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] 清 悦久, 田代 充, 山口健太郎ほか: "コールドスプレーイオン化質量分析法"分析化学. (印刷中).

    • Related Report
      2003 Annual Research Report

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Published: 2003-04-01   Modified: 2016-04-21  

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