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Study on cold and heat denaturation of amyloid fibrils of intrinsically disordered alpha-synuclein

Research Project

Project/Area Number 15K18518
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeMulti-year Fund
Research Field Biophysics
Research InstitutionOsaka University

Principal Investigator

YOUNG-HO LEE  大阪大学, たんぱく質研究所, 講師 (70589431)

Project Period (FY) 2015-04-01 – 2017-03-31
Project Status Completed (Fiscal Year 2016)
Budget Amount *help
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2016: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2015: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
KeywordsAlpha-synuclein / Amyloid fibril / Cold denaturation / Heat denaturation / Kinetics / Protein aggregation / Protein misfolding / Thermodynamics
Outline of Final Research Achievements

I examined cold and heat denaturation of amyloid fibrils of intrinsically disordered alpha-synuclein (aSN) prepared at various temperatures (298, 310, 323, 333, and 343 K). Despite differences in temperature for aSN amyloidogenesis, all fibrils showed similar core regions and secondary structures with the distinct width. As temperature increased from 298 to 383 K, all types of aSN fibrils heat-denatured to monomers from approximately 343 K. Decreasing temperature from 298 to 273 K caused cold denaturation of fibrils to monomers. Intriguingly, the stability of aSN fibrils depended on temperature for preparing fibrils. aSN fibrils generated at higher temperature showed higher stability at high and low temperature as well as slower denaturation kinetics than fibrils formed at lower temperature. Taken together, the stability of aSN amyloid fibrils depended on growing temperature of fibrils which produced differences in higher order structures of amyloid fibrils.

Report

(3 results)
  • 2016 Annual Research Report   Final Research Report ( PDF )
  • 2015 Research-status Report
  • Research Products

    (34 results)

All 2017 2016 2015 Other

All Int'l Joint Research (8 results) Journal Article (13 results) (of which Int'l Joint Research: 8 results,  Peer Reviewed: 13 results,  Open Access: 5 results,  Acknowledgement Compliant: 6 results) Presentation (7 results) (of which Invited: 7 results) Book (6 results)

  • [Int'l Joint Research] Eotvos Lorand University(Hungary)

    • Related Report
      2016 Annual Research Report
  • [Int'l Joint Research] University of Michigan(米国)

    • Related Report
      2016 Annual Research Report
  • [Int'l Joint Research] Univerisity of Leeds(英国)

    • Related Report
      2016 Annual Research Report
  • [Int'l Joint Research] Malvern(英国)

    • Related Report
      2016 Annual Research Report
  • [Int'l Joint Research] Eotvos Lorand University(Hungary)

    • Related Report
      2015 Research-status Report
  • [Int'l Joint Research] University of Michigan(米国)

    • Related Report
      2015 Research-status Report
  • [Int'l Joint Research] University of Leeds(英国)

    • Related Report
      2015 Research-status Report
  • [Int'l Joint Research] Malvern(英国)

    • Related Report
      2015 Research-status Report
  • [Journal Article] Kinetics and polymorphs of yeast prion Sup35NM amyloidogenesis International Journal of Biological Macromolecules2017

    • Author(s)
      Misaki Kinoshita, Yuxi Lin, Masatoshi Nakatsuji, Takashi Inui, Young-Ho Lee
    • Journal Title

      Int. J. Biol. Macromolec.

      Volume: -

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Mechanistic and structual basis of bioengineered bovine Cathelicidin-5 with optimized therapeutic activity2017

    • Author(s)
      Bikash R. Sahoo, Kenta Maruyama, Jyotheeswara R. Edula, Takahiro Tougan, Yuxi Lin, Young-Ho Lee, Toshihiro Horii, and Toshimichi Fujiwara
    • Journal Title

      Scientific Reports

      Volume: 7:44781 Issue: 1 Pages: 44781-44796

    • DOI

      10.1038/srep44781

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Practical Tips of Calorimetric Experiments using PEAQ-ITC2017

    • Author(s)
      Misaki Kinoshita, Ju Yaen Kim, Yuxi Lin, Young-Ho Lee
    • Journal Title

      Malvern Technical note

      Volume: -

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Biophysical Applications of Calorimetric Methods to Examination of Protein Misfolding and Aggregation2017

    • Author(s)
      Tatsuya Ikenoue, Misaki Kinoshita, Yuxi Lin, and Young-Ho Lee
    • Journal Title

      Malver white paper

      Volume: -

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed
  • [Journal Article] A. Reduced Lipid Bilayer Thickness Regulates the Aggregation and Cytotoxicity of Amyloid-β2017

    • Author(s)
      Kyle J. Korshavn, Cristina Satriano, Yuxi Lin, Rongchun Zhang, Mark Dulchavsky, Anirban Bhunia, Magdalena I. Ivanova, Young-Ho Lee, Carmelo La Rosa, Mi Hee Lim, Ayyalusamy Ramamoorthy
    • Journal Title

      J. Biol. Chem.

      Volume: 292 Issue: 11 Pages: 4638-4650

    • DOI

      10.1074/jbc.m116.764092

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Energetic basis on interactions between ferredoxin and ferredoxin NADP+ reductase at varying physiological conditions.2016

    • Author(s)
      Kinoshita, M., Kim, J.Y., S., Lin, Y., Hun Mok K., Kataoka, Y., Ishimori, K., Markova, N., Kurisu, G., Hase, T., Lee, Y.H.
    • Journal Title

      Biochem. Biophys. Res. Commun.

      Volume: 482 Issue: 4 Pages: 909-915

    • DOI

      10.1016/j.bbrc.2016.11.132

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Noncovalent Forces Tune the Electron Transfer Complex between Ferredoxin and Sulfite Reductase to Optimize Enzymatic Activity2016

    • Author(s)
      Ju Yaen Kim, Misaki Kinoshita, Satoshi Kume, Guy T. Hanke, Toshihiko Sugiki, John E. Ladbury, Chojiro Kojima, Takahisa Ikegami, Genji Kurisu, Yuji Goto, Toshiharu Hase, Young-Ho Lee
    • Journal Title

      Biochem J.

      Volume: 473 Pages: 3837-3854

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Solution NMR structure and inhibitory effect against amyloid-β fibrillation of Humanin containing a D-isomerized serine residue2016

    • Author(s)
      Nesreen Alsanousi, Toshihiko Sugiki, Kyoko Furuita, Masatomo So, Young-Ho Lee, Toshimichi Fujiwara, and Chojiro Kojima
    • Journal Title

      Biochem. Biophys. Res. Commun.

      Volume: 477 Issue: 4 Pages: 647-653

    • DOI

      10.1016/j.bbrc.2016.06.114

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Mutations affecting the internal equilibrium of the reaction catalyzed by 6-aminohexanoate-dimer hydrolase2016

    • Author(s)
      Negoro S, Kawashima Y, Shibata N, Kobayashi T, Baba T, Lee YH, Kamiya K, Shigeta Y, Nagai K, Takehara I, Kato D, Takeo M, Higuchi Y
    • Journal Title

      FEBS Lett

      Volume: 590 Issue: 18 Pages: 3133-3143

    • DOI

      10.1002/1873-3468.12354

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] A linker for the isopeptide mimetics by the peptide ligation2016

    • Author(s)
      Kawakami Toru, Mishima Yuichi, Kinoshita Misaki, Lee Young-Ho, Suetake Isao
    • Journal Title

      Tetrahedron Letters

      Volume: -

    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
  • [Journal Article] Amorphous Aggregation of Cytochrome c with Inherently Low Amyloidogenicity Is Characterized by the Metastability of Supersaturation and the Phase Diagram2016

    • Author(s)
      Lin Yuxi, Kardos József, Imai Mizue, Ikenoue Tatsuya, Kinoshita Misaki, Sugiki Toshihiko, Ishimori Koichiro, Goto Yuji, and Lee Young-Ho
    • Journal Title

      Langmuir

      Volume: 32 Issue: 8 Pages: 2010-2022

    • DOI

      10.1021/acs.langmuir.5b03810

    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
  • [Journal Article] X-ray Structure and Nuclear Magnetic Resonance Analysis of the Interaction Sites of the Ga-Substituted Cyanobacterial Ferredoxin2015

    • Author(s)
      Mutoh Risa, Muraki Norifumi, Shinmura Kanako, Kubota-Kawai Hisako, Lee Young-Ho, Marc M. Nowaczyk, Matthias Rögner, Hase Toshiharu, Ikegami Takahisa, Kurisu Genji
    • Journal Title

      Biochemistry

      Volume: 54 Issue: 39 Pages: 6052-6061

    • DOI

      10.1021/acs.biochem.5b00601

    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
  • [Journal Article] Physicochemical Nature of Interfaces Controlling Ferredoxin NADP+ Reductase Activity through Its Interprotein Interactions with Ferredoxin2015

    • Author(s)
      Kinoshita Misaki, Kim Ju Yaen, Kume Satoshi, Sakakibara Yukiko, Sugiki Toshihiko, Kojima Chojiro, Kurisu Genji, Ikegami Takahisa, Hase Toshiharu, Kimata-Ariga Yoko, Lee Young-Ho
    • Journal Title

      Biochimica et Biophysica Acta-Bioenergetics

      Volume: 1847 Issue: 10 Pages: 1200-1211

    • DOI

      10.1016/j.bbabio.2015.05.023

    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
  • [Presentation] Principle of protein misfolding and aggregation ‐ new findings of amyloid formation from thermodynamics2016

    • Author(s)
      Lee Young-Ho
    • Organizer
      RIKEN CLST invited seminar
    • Place of Presentation
      Kobe, Japan
    • Year and Date
      2016-03-18
    • Related Report
      2015 Research-status Report
    • Invited
  • [Presentation] The interaction of sulfite reductase with ferredoxin and its relation to enzyme activity2016

    • Author(s)
      Lee Young-Ho
    • Organizer
      Invited seminar in Hokkaido University
    • Place of Presentation
      Hokkaido, Japan
    • Year and Date
      2016-03-12
    • Related Report
      2015 Research-status Report
    • Invited
  • [Presentation] Thermodynamic study on interprotein interaction between oppositely-charged electron transfer proteins2016

    • Author(s)
      Lee Young-Ho
    • Organizer
      Institute for Protein Research seminar
    • Place of Presentation
      Osaka, Japan
    • Year and Date
      2016-02-02
    • Related Report
      2015 Research-status Report
    • Invited
  • [Presentation] New findings on thermodynamic properties of disease-inducing protein aggregation, Japan-Korea Bilateral Symposium on Multi-Scale Structural Biology2016

    • Author(s)
      Young-Ho Lee
    • Organizer
      International seminar at Institute for Protein Research
    • Place of Presentation
      大阪大学(大阪府吹田市)
    • Related Report
      2016 Annual Research Report
    • Invited
  • [Presentation] 1. 等温滴定熱量測定を用いた分子間相互作用の熱力学2016

    • Author(s)
      李 映昊
    • Organizer
      Malvern ウェブナー
    • Place of Presentation
      神戸国際ビジネスセンター(兵庫県、神戸市)
    • Related Report
      2016 Annual Research Report
    • Invited
  • [Presentation] The study on thermodynamic property of protein misfolding and aggregation2015

    • Author(s)
      Lee Young-Ho
    • Organizer
      第38回日本分子生物学会・第88回日本生化学会大会
    • Place of Presentation
      Kobe, Japan
    • Year and Date
      2015-12-01
    • Related Report
      2015 Research-status Report
    • Invited
  • [Presentation] Toward the establishment of thermodynamics on protein misfolding and aggregatio2015

    • Author(s)
      Lee Young-Ho
    • Organizer
      Open seminar in Osaka Prefecture University
    • Place of Presentation
      Osaka, Japan
    • Year and Date
      2015-04-22
    • Related Report
      2015 Research-status Report
    • Invited
  • [Book] Advanced Techniques to Detect Protein-Protein Interactions using Solution Nuclear Magnetic Resonance (Part II: Cross-saturation, Paramagnetic Effects, and In-cell NMR) Protein-Protein Interactions (PPIs): Types, Methods for Detection and Analysis2017

    • Author(s)
      Toshihiko Sugiki, Young-Ho Lee
    • Total Pages
      21
    • Publisher
      Nova Science Publishers, Inc.
    • Related Report
      2016 Annual Research Report
  • [Book] Advanced Techniques to Detect Protein-Protein Interactions using Solution Nuclear Magnetic Resonance (Part I: Chemical Shift Perturbation and Residual Dipolar Coupling) Protein-Protein Interactions (PPIs): Types, Methods for Detection and Analysis2017

    • Author(s)
      Young-Ho Lee, Toshihiko Sugiki
    • Total Pages
      21
    • Publisher
      Nova Science Publishers, Inc.
    • Related Report
      2016 Annual Research Report
  • [Book] Molecular Structure of Amyloid Fibrils Revealed by Thermodynamics Protein-Protein Interactions (PPIs): Types, Methods for Detection and Analysis2017

    • Author(s)
      Tatsuya Ikenoue, Yuxi Lin, Misaki Kinoshita, Kazumasa Sakurai, Ayyalusamy Ramamoorthy, Yuji Goto, Young-Ho Lee
    • Total Pages
      13
    • Publisher
      Nova Science Publishers, Inc.
    • Related Report
      2016 Annual Research Report
  • [Book] Thermodynamics of Protein-Protein Interactions Examined by Isothermal Titration Calorimetry Protein-Protein Interactions (PPIs): Types, Methods for Detection and Analysis2017

    • Author(s)
      Ju Yaen Kim, Misaki Kinoshita, Takashi Inui, Genji Kurisu, Yuji Goto, Toshiharu Hase, Koichiro Ishimori, Young-Ho Lee
    • Total Pages
      16
    • Publisher
      Nova Science Publishers, Inc.
    • Related Report
      2016 Annual Research Report
  • [Book] Methods in Molecular Biology: Biochemical and Biophysical Methods to Examine Effects of Site-directed Mutagenesis on Enzymatic Activities and Interprotein Interactions2017

    • Author(s)
      Misaki Kinoshita, Ju Yaen Kim, Yuxi Lin, Natalia Markova, Toshiharu Hase, Young-Ho Lee
    • Total Pages
      22
    • Publisher
      Springer Nature
    • Related Report
      2016 Annual Research Report
  • [Book] Methods in Molecular Biology (In vitro mutagenesis: Methods and Protocol)2016

    • Author(s)
      Kinoshita Misaki, Kim Ju Yaen, Lin Yuxi, Natalia Markova, Toshiharu Hase, Lee Young-Ho
    • Total Pages
      500
    • Publisher
      Springer Science+Business Media
    • Related Report
      2015 Research-status Report

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Published: 2015-04-16   Modified: 2022-02-16  

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