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Development of solid NMR methods for structural determination of a peptide

Research Project

Project/Area Number 13558081
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Structural biochemistry
Research InstitutionKYOTO UNIVERSITY

Principal Investigator

TERAO Takehiko  Kyoto Univ., Graduate School of Sci., Prof., 大学院・理学研究科, 教授 (50093274)

Co-Investigator(Kenkyū-buntansha) SUGISAWA Hisashi  JEOL, Application & Research Center, Analytical Instruments Division, Researcher, 分析機器技術本部・応用研究センター, 研究員
TAKEGOSHI Kiyonori  Kyoto Univ., Graduate School of Sci., Asso. Prof., 大学院・理学研究科, 助教授 (10206964)
Project Period (FY) 2001 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥9,000,000 (Direct Cost: ¥9,000,000)
Fiscal Year 2002: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 2001: ¥6,500,000 (Direct Cost: ¥6,500,000)
Keywordssolid NMR / structural determination / peptide / dihedral angle / nuclear distance measurements
Research Abstract

The aim of this research is to establish a solid-NMR method, which enables us to determine a 3D structure of a fully ^<13>C-labeled biomolecule. In the first year, we developed a new method and modified our NMR machine as follows:
(1) Development of ^1H-^<13>C dipolar assisted rotational resonance (DARR)
We developed a method enabling us to determine a large number of distances by one 2D experiment. Such method has long been desired but not yet been realized. Since DARR deos not apply rf irradiation to the observed spins, the mixing time for distance measurement is limited by generally long spin-lattice relaxation times. This enables us to measure longer distances. Further, we showed that DARR has many features suitable for application to a multiply isotope-labeled sample, such as, observation is done unedr high-resolition conditions.
(2) Installation of low-temperature observation apparatus
For acculate determination of a molecular structure, suppression of local molecular motion by lower … More ing the sample temperature is prerequisite. We bought a recently developed low temperature NMR observation system and also a liquid-nitrogen recondensation system. These systems were combined with our NMR system to realize safe and stable long low-temperature experiments.
In the second year, we applied the DARR method to determine the structure of the eledoisin-related peptide which is a neurotransmitter. A fully ^<13>C and ^<15>N labeled peptide sample conposed of six residues (K-F-I-G-L-M), which is the common C-terminal sequence of the tachykinin family, was synthesized by using a solid synthesis procedure. The structural determination was done as follows, 81 relevant C-C distances were obtaned from the 2D ^<13>C-^<13>C distance-correlation experiment with DARR. These distances were fed into the X-Plor program developed at Yale University and a 3D structure was successfully obtaned based on the 81 distance constraines. Further, the four φ dihedral angles of the peptide main chain were determined and fed also into the X-Plor program as angle constrains. It was shown that the precision of the structure becomes better by adding these dihedral angle constraints. To summarize, in this project, we can establish the solid-NMR method enabling us to determine a 3D structure of a peptide molecule with reasonable acculacy by a few numbers of 2D solid-NMR experiments using one multiply labeled peptide sample. Less

Report

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

    (24 results)

All Other

All Publications (24 results)

  • [Publications] K.Takegoshi: "^<13>C-^1H dipolar-assisted rotational resonance in magic angle spinning NMR"Chem. Phys. Lett.. 344(5-6). 631-637 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] K.Takegoshi: "^<13>C nuclear Overhauser polarization nuclear magnetic resonance in rotating solids : Replacement of cross polarization in uniformly ^<13>C labeled molecules with methyl groups"J. Chem. Phys.. 117(4). 1700-1707 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] K.Takeda: "Zero-field electron spin resonance and theoretical studies of light penetration into single crystal and polycrystalline material doped with molecules photoexcitable to the triplet state via intersystem crossing"J. Chem. Phys.. 117(10). 4940-4946 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Y.Nishiyama: "γ-Encoding transform NMR for uniaxially oriented samples"Chem. Phys. Lett.. 352(5-6). 479-485 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T.Miyoshi: "Effects of Xe Gas on Segmental Motion in a Polymer Blend As Studied by ^<13>C and ^<129>Xe High-Pressure MAS NMR"Macromolecules. 35(1). 151-154 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] K.Takegoshi: "^<13>C-^1H dipolar-driven ^<13>C-^<13>C recoupling without ^<13>C rf irradiation in nuclear magnetic"J. Chem. Phys.. 118(5). 2325-2341 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] K. Takegoshi et. al.: "^<13>C-^1H dipolar-assisted rotational resonance in magic angle spinning NMR"Chem. Phys. Lett.. 344(5-6). 631-637 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] K. Takegoshi and T. Terao: "^<13>C nuclear Overhauser polarization nuclear magnetic resonance in rotating solids: Replacement of cross polarization in uniformly ^<13>C labeled molecules with methyl groups"J. Chem. Phys.. 117(4). 1700-1707 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] K. Takeda et. al.: "Zero-field electron spin resonance and theoretical studies of light penetration into single crystal and polycrystalline material doped with molecules photoexcitable to the triplet state via intersystem crossing"J. Chem. Phys.. 117(10). 4940-4946 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Y. Nishiyama and T. Terao: "γEncoding transform NMR for uniaxially oriented samples"Chem. Phys. Lett.. 352(5-6). 479-485 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T. Miyoshi et. al.: "Effects of Xe Gas on Segmental Motion in a Polymer Blend As Studied by ^<13>C and ^<129>Xe High-Pressure MAS NMR"Macromolecules. 35(1). 151-154 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] K. Takegoshi et. al.: "^<13>C-^1H dipolar-driven ^<13>C-^<13>C recoupling without ^<13>C rf irradiation in nuclear magnetic resonance of rotating solids"J. Chem. Phys.. 118(5). 2325-2341 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Y.Nishiyama: "γ-Encoding transform NMR for uniaxially oriented samples"Chem.Phys.Lett.. 352(5-6). 479-485 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] K.Takegoshi: "^<13>Cnuclear Overhauser polarization nuclear magnetic resonance in rotating solids : Replacement of cross polarization in uniformly ^<13>C labeled molecules with methyl groups"J.Chem.Phys.. 117(4). 1700-1707 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] K.Takeda: "Zero-field electron spin resonance and theoretical studies of light penetration into single crystal and polycryslalline material doped with molecules photoexcitable to the triplet state via intersystem crossing"J.Chem.Phys.. 117(10). 4940-4946 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Y.Nishiyama: "^<13>C NMR spectral assignments for nematic liquid crystals by 2D chemical shift γ-encoding NMR"J.Magn.Reson.. 158(1/2). 60-64 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] T.Miyoshi: "Effects of Xe Gas on Segmental Motion in a Polymer Blend As Studied by ^<13>C and ^<129>Xe High-Pressure MAS NMR"Macromolecules. 35(1). 151-154 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] K.Takegoshi: "^<13>C-^1H dipolar-driven ^<13>C-^<13>C recoupling without ^<13>C rf irradiation in nuclear magnetic resonance of rotating solids"J.Chem.Phys.. 118(5). 2325-2341 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] K.Takegoshi: "^1H decoupling with third averaging in solid NMR"Chem. Phys. Lett.. 341(5-6). 540-544 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] K.Takegoshi: "^<13>C-^1H dipolar-assisted rotational resonance in magic angle spinning NMR"Chem. Phys. Lett.. 344(5-6). 631-637 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] K.Takeda: "Dynamic nuclear polarization by photoexcited-triplet electron spins in polycrystalline samples"Chem. Phys. Lett.. 345(1-2). 166-170 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] K.Takegoshi: "Indirect High-Resolution Observation of ^<14>N NMR in Rotating Solids"J. Am. Chem. Soc.. 123(43). 10786-10787 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] A.Asano: "Free Volume Study of Amorphous Polymers Detected by Solid State ^<13>C NMR Linewidth Experiments"J. Chem. Phys.. 115(18). 8665-8669 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] T.Miyoshi: "Effects of Xe Gas on Segmental Motion in a Polymer Blend As Studied by ^<13>C and ^<129>Xe High-Pressure MAS NMR"Macromolecules. 35(1). 151-154 (2002)

    • Related Report
      2001 Annual Research Report

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

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