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1998 Fiscal Year Final Research Report Summary

NMR Analysis of Pressure-Induced Structural Changes in Prioteins

Research Project

Project/Area Number 09480177
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Biophysics
Research InstitutionKOBE UNIVERSITY

Principal Investigator

AKASAKA Kazuyuki  Kobe University, Graduate School of Science and Technology, Professor, 大学院・自然科学研究科, 教授 (50025368)

Co-Investigator(Kenkyū-buntansha) TAMURA Atuo  Kobe, University Graduate School of Science and Technology Lecturer, 大学院・自然科学研究科, 講師 (90273797)
YAMADA Hiroaki  Kobe Univdersity, Faculty of Sciences, Asssociate Professor, 理学部, 助教授 (90030767)
Project Period (FY) 1997 – 1998
Keywordshigh pressure NMR / protein structure / 15N chemical shift / hydrogen bond / conformational change / NOE / BPTI / local unfolding
Research Abstract

1. We utilized a novel high pressure NMR technique that was developed in our laboratory to study the effect of pressure on protein structures in solution. The technique works on proteins in solution at any pressure between 1 and 2000 bar on a modern high field NMR spectrometer operating at 17.6 T or 750 MHz for 1H.
2. We applied this technique to a number of proteins including 15N -uniformly labeled BPTI (basic pancreatic trypsin inhibitor), helix (1-36) of Bacteriorhodopsin, HPr (histidine-containing phospho-carrier protein), RalGEF-RBD (Ral guanine-nucleotide exchange factor-Ras binding domain). Pressure-induced 1H and 15N chemical shift changes revealed in 15N /1H two-dimensional spectra of peptide NH groups showed that the entire backbone structure of a folded protein responds to pressure, but that the response is non-uniform over the sequence.
3. From pressure-induced chemical shifts, we conclude (1) Distances of practically all the hydrogen bonds of amide NH groups, either with carbonyls or solvent water, are shortened by pressure ; (2) Pressure-induced 15N shifts report non-uniform conformational changes in the polypeptide backbone by pressure.
4. From pressure-induced changes in NOE (Nuclear Overhauser Effect) intensities for BPTI, we conclude that pressure induces selective compactions of certain regions of the protein, with concomitant slowing down of flip-flop motions of some aromatic rings.
5. Finally, from signal intensity measurements in RalGEF-RBD, we found that pressure induces local unfolding of the protein preferentially in loop regions above 1000 bar, followed by unfolding of the entire protein at 2000 bar.

  • Research Products

    (13 results)

All Other

All Publications (13 results)

  • [Publications] K.Akasaka: "Pressure-induced changes in the folded structure of lysozyme" J.Mol.Biol.271. 671-678 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] K.Inoue: "High pressure NMR study of a small protein gurmarin" J.Biomol.NMR. 10. 535-541 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] H.Li: "Effect of pressure on individual hydrogen in proteins.Basic pancreatic trypsin inhibitor" Biochemistry. 37. 1167-1173 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 赤坂 一之: "高磁場高圧NMRが開く新しい世界、蛋白質の立体構造変化と揺らぎを鋭敏に観測" 化学と生物. 36巻・7号. 423-425 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] K.Akasaka: "Bilogical NMR Spectroscopy,Chapter 12" Oxford University Press, 12 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 赤坂 一之: "高圧バイオテクノロジー 第2章" さんえい出版, 5 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] K.Akasaka: "Trends/Graduate School of Science and Technology, Kobe University 1999" Spring-Verlag, 12 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] K.Akasaka: "Pressure-induced changes in the folded structure of lysozyme" J.Mol.Biol.Vol.271. 671-678 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K.Inoue: "High pressure NMR study of a small prote in gurmarin" J.Biomol.NMR. Vol.10. 535-541 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] H.Li: "Effect of pressure on individual hydrogen bonds in proteins, Basic pancreatic trypsin inhibitor." Biochemistry. Vol.37. 1167-1173 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K.Akasaka: "The new world opened by high field-high pressure NMR spectroscopy" Chemistry and Biology. Vol.36, No.7. 423-425 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K.Akasaka: "NMR Approaches to the Heat-, Cold-and Pressure-Induced Unfolding of Proteins." Biological NMR Spectroscopy, Oxford University Press, Chapt. 12. 12 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K.Akasaka: New insight into protein dynamics from high pressure NMR at high magnetic field. High pressure biotechnology, Chapt.2. Sanei Publishers, 5 (1998)

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

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Published: 1999-12-08  

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