Characterization of the excited state of prion protein using high-pressure NMR technique

• Project/Area Number: 22770158
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Biophysics
• Research Institution: Kinki University
• Principal Investigator: MAENO Akihiro 近畿大学, 先端技術総合研究所, 研究員 (70570951)
• Project Period (FY): 2010 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2012: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000); Fiscal Year 2011: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000); Fiscal Year 2010: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
• Keywords: プリオン蛋白質 / 核磁気共鳴 / 圧力 / 励起構造

Research Abstract

The purpose of this study is to clarify the conformational basis of infection mechanism and to characterize the excited state of prion protein which causes a series of prion diseases defined by prion hypothesis. We established the protocols for the high-level expression and the purification of 15N-labeled full length hamster prion. In the experiment of NMR transverse relaxation dispersion that expected to enable the quantitative analysis of the excited state which is stabilized under pressure, the typical CPMG profiles have not been obtained and those experimental conditions have failed to be optimized. These results suggest that conformational fluctuations between the basic folded state and the excited state for haPrPC may take place beyond detection range (milli-second time scale) of NMR R2 dispersion measurement.

Research Products

• [Journal Article] Extensively hydrated but folded : A novel state of globular proteins stabilized at high pressure and low temperature (2012)
  • Author(s): Sunilkumar P.N., Maeno A, et al
  • Journal Title: Biophysical Journal Volume: 102 Issue: 2 Pages: L8-L10
  • DOI: 10.1016/j.bpj.2011.12.015
  • Peer Reviewed
• [Journal Article] Pressure-Accelerated Dissociation of Amyloid Fibrils in Wild-Type Hen Lysozyme (2012)
  • Author(s): Budda R.Shah, 他
  • Journal Title: Biophysical Journal Volume: 102 Issue: 1 Pages: 121-126
  • DOI: 10.1016/j.bpj.2011.10.041
  • Peer Reviewed
• [Journal Article] Pressure-accelerated dissociation of amyloid fibrils in wild-type hen lysozyme. (2011)
  • Author(s): Buddha R Shah, Akihiro Maeno, Hiroshi Matsuo, Hideki Tachibana and Kazuyuki Akasaka
  • Journal Title: Biophys. J. Volume: 102(1) Pages: 121-126
  • Peer Reviewed
• [Presentation] Crucial roles of cavity in the conformational fluctuation of a protein. (2012)
  • Author(s): 前野覚大
  • Organizer: 第51回 NMR討論会
  • Place of Presentation: ウインクあいち(愛知県)
  • Year and Date: 2012-11-09
• [Presentation] Crucial roles of cavity in the conformational fluctuation of a protein. (2012)
  • Author(s): 前野覚大
  • Organizer: 7th International Conference on High Pressure Bioscience and Biotechnology
  • Place of Presentation: ピアザ淡海(滋賀県)
  • Year and Date: 2012-10-29
• [Presentation] Single mutation on a loop alters the key dynamics of the core, but not the average structure. (2012)
  • Author(s): 前野覚大
  • Organizer: 第50回日本生物物理学会年会
  • Place of Presentation: 名古屋大学(愛知県)
  • Year and Date: 2012-09-23
• [Presentation] Single mutation on a loop alters the key dynamics of the core, but not the average structure (2012)
  • Author(s): 前野 覚大
  • Organizer: 第50回日本生物物理学会年会
  • Place of Presentation: 名古屋大学（愛知県）
• [Presentation] Crucial roles of cavity in the conformational fluctuation of a protein (2012)
  • Author(s): 前野 覚大
  • Organizer: 7th International Conference on High Pressure Bioscience and Biotechnology (HPBB2012)
  • Place of Presentation: ピアザ淡海（滋賀県）
• [Presentation] Crucial roles of cavity in the conformational fluctuation of a protein (2012)
  • Author(s): 前野 覚大
  • Organizer: 第51回NMR討論会
  • Place of Presentation: ウインクあいち（愛知県）
• [Presentation] Cavity hydration and the dynamics of a protein. (2011)
  • Author(s): 前野覚大
  • Organizer: 6th International Meeting on Biomolecules under Pressure
  • Place of Presentation: 大津市旧公会堂(滋賀県)
  • Year and Date: 2011-12-15
• [Presentation] Cavity hydration and the dynamics of a protein (2011)
  • Author(s): 前野覚大
  • Organizer: IMBP2011
  • Place of Presentation: 旧大津公会堂(大津)
  • Year and Date: 2011-12-12
• [Presentation] High pressure NMR study of DHFR from a deep-sea bacterium. (2011)
  • Author(s): 前野覚大
  • Organizer: International Symposium on Nuclear Magnetic Resonance 2011
  • Place of Presentation: 大桟橋ホール(神奈川県)
  • Year and Date: 2011-11-17
• [Presentation] High pressure NMR study of DHFR from a deep-sea bacterium (2011)
  • Author(s): 前野覚大
  • Organizer: ISNMR2011
  • Place of Presentation: 大桟橋ホール(横浜)
  • Year and Date: 2011-11-15
• [Presentation] Cavity hydration and the dynamics of a protein (2011)
  • Author(s): 前野覚大
  • Organizer: 第49回生物物理学会年会シンポジウム(主催者)
  • Place of Presentation: 兵庫県立大学(姫路)
  • Year and Date: 2011-09-17
• [Presentation] Cavity hydration and the dynamics of a protein. (2011)
  • Author(s): 前野覚大
  • Organizer: 第49回生物物理学会年会シンポジウム
  • Place of Presentation: 兵庫県立大学(兵庫県)
  • Year and Date: 2011-09-16
• [Presentation] Cavity Hydration: A Gate Way to the High-Energy Paradigm? (2010)
  • Author(s): 前野覚大
  • Organizer: 第48回生物物理学会年会シンポジウム
  • Place of Presentation: 東北大学(宮城県)
  • Year and Date: 2010-09-21
• [Presentation] Cavity Hydration : A Gate Way to the High-Energy Paradigm? (2010)
  • Author(s): 前野覚大
  • Organizer: 第48回生物物理学会年会
  • Place of Presentation: 東北大学川内キャンパス(宮城県)
  • Year and Date: 2010-09-21
• [Presentation] Conformational fluctuation of highly stable form of staphylococcal nuclease Δ+P HS and its V66K variant studied by high pressure NMR spectroscopy (2010)
  • Author(s): 前野覚大
  • Organizer: ICMRBS2010
  • Place of Presentation: Cairns Convention Center (Australia)
  • Year and Date: 2010-08-25
• [Presentation] Conformational fluctuation of highly stable form of staphylococcal nuclease .+PHS and its V66K variant studied by high pressure NMR spectroscopy. (2010)
  • Author(s): Maeno A, et al.
  • Organizer: 24thInternational Conference on Magnetic Resonance in Biological Systems
  • Place of Presentation: ケアンズ(オーストラリア)
  • Year and Date: 2010-08-23