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1999 Fiscal Year Final Research Report Summary

Development of Solid-State NMR Method for Studying Higher-Order Structure of Proteins

Research Project

Project/Area Number 09558091
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Biophysics
Research InstitutionYokohama National University

Principal Investigator

FUJIWARA Toshimichi  Yokohama National University, Faculty of Engineering, Associate Professor, 工学部, 助教授 (20242381)

Co-Investigator(Kenkyū-buntansha) HINOMOYO Tetsu  JOEL Ltd., Division of Analytical Instruments, Vice-Director, 分析機器技術本部, 統括次長(研究職)
AKUTSU Hideo  Yokohama National University, Faculty of Engineering, Professor, 工学部, 教授 (60029965)
Project Period (FY) 1997 – 1999
KeywordsSolid-State NMR / Labeling with Stable Isotope / Analysis of Protein Higher-Order Structure / Structural Biology
Research Abstract

Multi-dimensional correlation NMR has been developed especially for ィイD113ィエD1C, ィイD115ィエD1N labeled biomolecules in solid states. Multi-dimensional NMR that has two or three isotropic shift evolution periods under magic-angle spinning conditions resolve ィイD113ィエD1C and ィイD115ィエD1N signals of uniformly isotope-labeled biomolecules. Dipolar couplings for ィイD113ィエD1C and ィイD115ィエD1 spins in labeled molecules provide spin connectivity for signal assignment and internuclear distances information for the structure determination. Analysis of the mutual orientation of anisotropic interactions for ィイD113ィエD1C and ィイD115ィエD1N spins such as chemical shift anisotropies and dipolar interactions permit the determination of dihedral angles.
Dihedral angles can be determined from the correlations between anisotropic interactions of different spins, because such correlation spectra depend on mutual orientations of anisotropic interactions which specify dihedral angles. These correlation spectra were ob … More tained with experiments in which evolutions of anisotropic interactions are connected by a dipolar mixing period. This experiment is combined with the isotropic-shift correlation to enhance the spectral resolution. Example are shown for the correlation of the ィイD113ィエD1CィイD11ィエD1H and ィイD113ィエD1CィイD113ィエD1C dipolar couplings in an ィイD11ィエD1HィイD1α13ィエD1CィイD1αィエD1-ィイD113ィエD1CィイD1β1ィエD1HィイD1βィエD1 moiety and for the correlation of the ィイD113ィエD1CO chemical shift anisotropy and CH dipolar interactions. The experimental pulse sequence includes ィイD113ィエD1CィイD11ィエD1H dipolar evolution periods for ィイD113ィエD1CィイD1αィエD1 and ィイD113ィエD1CィイD1βィエD1H, which are correlated with a coherent ィイD113ィエD1CィイD1α13ィエD1CィイD1βィエD1 dipolar mixing period. This method enables the experimental determination of all the structural parameters for the four-spin system.
A pulse sequence applied synchronously with sample-spinning is designed for enhancing magnetization transfer between ィイD113ィエD1C spins. This method improve the transfer efficiency by reducing γ-dependence of the zero-quantum dipolar mixing operator. This efficiency was verified for fully ィイD113ィエD1C labeled alanine.
Sample preparation for solid state NMR has also been improved. Rapid sample cooling and cooling with protectant such as trehalose reduce the signal broadening due to distribution of molecular conformations. This method was applied to ATP complexed with FィイD21ィエD2-ATP synthase β subunit. The conformation was monitored with ィイD131ィエD1P-NMR. Less

  • Research Products

    (16 results)

All Other

All Publications (16 results)

  • [Publications] Toshimichi Fujiwara: "Multidimensional Solid-State Nuclear Magnetic Resonance for Correlating Anisotropic Interactionss under Magic-Angle Spinning Conditions"J. Magn. Reson.. 124. 147-153 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Suhkman Kim: "The Interactions of Ferric and Ferrous Cytochrome c with Cardiolipin in Phospholipid Membranes Studied by Solid-State ^2H and ^<31>P NMR"J. Mol. Struct.. 441. 183-188 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tomoaki Ohmura: "Paramagnetic Inversion of the Sign of the Interference Contribution to the Transverse Relaxation of the Imido Protons of the Coordinated Imidazoles in the Uniformly ^<15>N Labeled Cytochrome c_3"J. Magn. Reson.. 131. 367-372 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Toshimichi Fujiwara: "Multidimensional Solid-State Nuclear Magnetic Resonance for Determining the Dihedral Angel from the Correlation of ^<13>C-^1H and ^<13>C-^<13>C Dipolar Interactions under Magic-Angle Spinning Conditions"J. Chem. Phys.. 109. 2380-2393 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Suhkman Kim: "Osmotic Stability of Muramyl Dipeptide-Bearing Liposomes and Molecular Miscibility in Their Membrances"Bull. Chem. Soc. Jpn.. 72. 541-548 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sumie Yoshioka: "Molecular Mobility of Protein in Lyophilized Formulations Linked to the Molecular Mobility of Polymer Excipients, as Determined by High Resolution ^<13>C Solid-State NMR"Pharmaceutical Research. 16. 1621-1625 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Toshimichi Fujiwara: "Compound Radio-Frequency Driven Recoupling Pulse Sequences for Efficient Magnetization Transfer by Homonuclear Dipolar Interaction under Magic Angle spinning Conditions"J. Magn. Reson.. (in press).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 阿久津秀雄: "構造生物学とその解析法(の「固体NMRとその利用」)"共立出版. 13 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 藤原敏道: "蛋白質・核酸・酸素 44(4)(の「固体核磁気共鳴法」)"共立出版. 8 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Toshimichi Fujiwara: "Multidimensional Solid-State Nuclear Magnetic Resonance for Correlating Anisotropic Interactions under Magic-angle Spinning Conditions"J. Magn. Reson.. 124. 147-153 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Suhkman Kim: "The Interactions of Ferric and Ferrous Cytochrome c with Cardiolipin in Phospholipid Membranes Studied by Solid-State ィイD12ィエD1H and ィイD131ィエD1P NMR"J. Mol. Struct.. 441. 147-153 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tomoaki Ohmura: "Paramagnetic Inversion of the Sign of the Interference Contribution to the Transverse Relaxation of the Imido Protons of the Coordinated Imidazoles in the Uniformly ィイD115ィエD1N Labeled Cytochrome cィイD23ィエD2"J. Magn. Reson.. 131. 367-372 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Toshimichi Fujiwara: "Multidimensional Solid-State Nuclear Magnetic Resonance for Determining the Dihedral Angle from the Correlation of ィイD113ィエD1C-ィイD11ィエD1H and ィイD113ィエD1C-ィイD113ィエD1H Dipolar Interactions under Magic-Angle Spinning Conditions"J. Chem. Phys.. 109. 2380-2393 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Suhkman Kim: "Osmotic Stability of Muramyl Dipeptide-Bearing Liposomes and Molecular Miscibility in Their Membranes"Bull. Chem. Soc. Jpns.. 72. 541-548 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sumie Yoshioka: "Molecular Mobility of Protein in Lyophilized Formation Linked to the Molecular Mobility of Polymer Excipients, as Determined by High Resolution ィイD113ィエD1C Solid-State NMR"Pharmaceutical Research. 16. 1621-1625 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Toshimichi Fujiwara: "Compound Radio-Frequency Driven Recoupling Pulse Sequences for Efficient Magnetization Transfer by Homonuclear Dipolar Interaction under Magic Angle Spinning Conditions"J. Magn. Reson.. (in press).

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2001-10-23  

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