Study on effect of electrostatic interactions on stability of electron transfer complexes and amyloid fibrils

• Project/Area Number: 25870407
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Structural biochemistry; Biophysics
• Research Institution: Osaka University
• Principal Investigator: LEE Young-Ho 大阪大学, たんぱく質研究所, 講師 (70589431)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2014: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2013: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: Amyloid fibril / Binding thermodynamics / Calorimetry / Electron transfer / Electrostatics / Enzyme activity / Interprotein interaction / Protein misfolding / Charge interaction / Enthalpy and entropy / Hydrophobic effect / Intermolecular force / Protein aggregation / Thermodynamics

Outline of Final Research Achievements

We have conducted thermodynamic studies on binding reactions between ferredoxin (Fd) and sulfite reductase (SiR) as well as Fd and ferredoxin NADP(+)-reductase (FNR) using calorimetry, NMR spectroscopy, and X-ray crystallographic analysis with FNR and SiR activity assays.
We have revealed that oppositely-charged Fd and SiR formed the electron transfer complex using attractive charge-charge interactions, and that non-electrostatic interactions was also important for fine tuning of binding modes between Fd and SiR to optimize SiR activity. The same results were also obtained in the binding system between Fd and FNR. We have also revealed that amyloid fibrils of alpha-synuclein cold-denatured below 293K due to the repulsive electrostatic interactions at low temperature.
We have clearly demonstrated that interprotein electrostatic interactions play an important roles in normal protein functions and in clearance of disease-related amyloid fibrils.

Research Products

• [Journal Article] Accurate Secondary Structure Prediction and Fold Recognition for Circular Dichroism Spectroscopy (2015)
  • Author(s): Andras Micsonai, Frank Wien, Linda Kernya, Young-Ho Lee, Yuji Goto, Matthieu Refregiers, and Jozsef Kardos
  • Journal Title: Proceedings of the National Academy of Sciences of the United States of America Volume: ー
  • Peer Reviewed
• [Journal Article] Optimized Co-Solute Paramagnetic Relaxation Enhancement for the Rapid NMR Analysis of a Highly Fibrillogenic Peptide (2015)
  • Author(s): Nur Alia Oktaviani, Michael W. Risor, Young-Ho Lee, Rik P. Megens, Djurre H. de Jong, Renee Otten, Ruud M. Scheek, Jan J. Enghild, Niels Chr. Nielsen, Takahisa Ikegami, and Frans A. A. Mulder
  • Journal Title: Journal of Biomolecular NMR Volume: ー
  • Peer Reviewed
• [Journal Article] Effects of a Reduced Disulfide Bond on Aggregation Properties of the Human IgG1 CH3 Domain (2015)
  • Author(s): Kazumasa Sakurai, Ryosuke Nakahata, Young-Ho Lee, Jozsef Kardos, Takahisa Ikegami, and Yuji Goto
  • Journal Title: Biochimica et Biophysica Acta Volume: ー Issue: 10 Pages: 1526-1535
  • DOI: 10.1016/j.bbapap.2015.02.020
  • Peer Reviewed
• [Journal Article] Ultrasonication-dependent formation and degradation of α-synuclein amyloid fibrils (2015)
  • Author(s): Hisashi Yagi, Aiko Mizuno, Masatomo So, Miki Hirano, Masayuki Adachi, Yoko Akazawa-Ogawa, Yoshihisa Hagihara, Tatsuya Ikenoue, Young-Ho Lee, Yasushi Kawata, Yuji Goto
  • Journal Title: Biochimica et Biophysica Acta Volume: 1854 Issue: 3 Pages: 209-217
  • DOI: 10.1016/j.bbapap.2014.12.014
  • Peer Reviewed
• [Journal Article] Small Liposomes Accelerate the Fibrillation of Amyloid β (2015)
  • Author(s): Mayu S. Terakawa, Hisashi Yagi, Masayuki Adachi, Young-Ho Lee, Yuji Goto
  • Journal Title: The Journal of Biological Chemistry Volume: 290 Issue: 2 Pages: 815-826
  • DOI: 10.1074/jbc.m114.592527
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Supersaturation-limited Amyloid Fibrillation of Insulin Revealed by Ultrasonication (2014)
  • Author(s): Hiroya Muta, Young-Ho Lee, Jozsef Kardos, Yuxi Lin, Hisashi Yagi, Yuji Goto
  • Journal Title: The Journal of Biological Chemistry Volume: 289 Issue: 26 Pages: 18228-18238
  • DOI: 10.1074/jbc.m114.566950
  • Peer Reviewed
• [Journal Article] Cold denaturation of alpha-synuclein amyloid fibrils. (2014)
  • Author(s): Ikenoue, T. , Lee, Y.-H. , Kardos, J., Sakai, M., Yagi, H., Kawata, Y. and Goto, Y.
  • Journal Title: Angew. Chem. Int. Ed. Volume: 53 Issue: 30 Pages: 7799-7804
  • DOI: 10.1002/anie.201403815
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] NMR Characterization of the Interaction of the Endonuclease Domain of MutL with Divalent Metal Ions and ATP (2014)
  • Author(s): Mizushima R, Kim JK, Suetake I, Tanaka H, Takai T, Kamiya N, Takano Y, Mishima Y, Tajima S, Goto Y, Fukui K, *Lee YH
  • Journal Title: PLoS One Volume: 9 Issue: 6 Pages: e98554-e98554
  • DOI: 10.1371/journal.pone.0098554
  • NAID: 120006557621
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Heat-induced Irreversible Denaturation of the Camelid Single Domain VHH Antibody is Governed by Chemical Modifications (2014)
  • Author(s): Yoko Akazawa-Ogawa, Mizuki Takashima, Young-Ho Lee, Takahisa Ikegami, Yuji Goto, Koichi Uegaki, and Yoshihisa Hagihara
  • Journal Title: Journal of Biological Chemistry Volume: 289 Issue: 22 Pages: 15666-15679
  • DOI: 10.1074/jbc.m113.534222
  • Peer Reviewed
• [Journal Article] Heat of supersaturation-limited amyloid burst directly monitored by isothermal titration calorimetry. (2014)
  • Author(s): Ikenoue, T., Lee, Y.-H., Kardos, J., Yagi, H., Ikegami, T., Naiki, H. and Goto, Y.
  • Journal Title: Proc. Natl. Acad. Sci. USA Volume: 111 Issue: 18 Pages: 6654-6659
  • DOI: 10.1073/pnas.1322602111
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Solubility and supersaturation-dependent protein misfolding revealed by ultrasonication. (2014)
  • Author(s): Lin, Y., Lee, Y.-H., Yoshimura, Y., Yagi, Y. and Goto, Y.
  • Journal Title: Langmuir Volume: 30 Issue: 7 Pages: 1845-1854
  • DOI: 10.1021/la403100h
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Fine-tuned broad binding capability of human lipocalin-type prostaglandin D synthase for various small lipophilic ligands. (2014)
  • Author(s): Kume S, Lee YH, Nakatsuji M, Teraoka Y, Yamaguchi K, Goto Y, Inui T
  • Journal Title: FEBS Lett Volume: 588 Issue: 6 Pages: 962-969
  • DOI: 10.1016/j.febslet.2014.02.001
  • Peer Reviewed
• [Journal Article] Heat of supersaturation-limited amyloid burst directly monitored by isothermal titration calorimetry (2014)
  • Author(s): Tatsuya Ikenouea(a)*, Young-Ho Lee(a)*, Jozsef Kardos, Hisashi Yagi, Takahisa Ikegami, Hironobu Naiki, and Yuji Goto (*equal contribution)
  • Journal Title: Proceedings of the National Academy of Sciences of the United States of America Volume: -
  • Peer Reviewed
• [Journal Article] Heat-Induced Irreversible Denaturation of the Camelid Single Domain VHH Antibody is Governed by Chemical Modifications (2014)
  • Author(s): Yoko Akazawa-Ogawa(a), Mizuki Takashima, Young-Ho Lee(b), Takahisa Ikegami, Yuji Goto, Koichi Uegaki, and Yoshihisa Hagihara
  • Journal Title: Journal of biological chemistry Volume: -
  • Peer Reviewed
• [Journal Article] Remaining structures at the N- and C-terminal regions of alpha-synuclein accurately elucidated by amide-proton exchange NMR with fitting (2013)
  • Author(s): Okazaki, H., Ohori, Y., Komoto, M., Lee, Y-H, Goto, Y., Tochio, N., and Nishimura, C.*
  • Journal Title: FEBS Letters Volume: 587 Issue: 22 Pages: 3709-3714
  • DOI: 10.1016/j.febslet.2013.09.039
  • Peer Reviewed
• [Journal Article] Structure, folding dynamics and amyloidogenesis of Asp76Asn β2-microglobulin: roles of shear flow, hydrophobic surfaces and α crystallin. (2013)
  • Author(s): Mangione PP, Esposito G, Relini A, Raimondi S, Porcari R, Giorgetti S, Corazza A, Fogolari F, Penco A, Goto Y, Lee YH, Yagi H, Cecconi C, Naqvi MM, Gillmore JD, Hawkins PN, Chiti F, Rolandi R, Taylor GW, Pepys MB, Stoppini M, Bellotti V.
  • Journal Title: J. Biol. Chem. Volume: 288 Issue: 43 Pages: 30917-30930
  • DOI: 10.1074/jbc.m113.498857
  • Peer Reviewed
• [Journal Article] Concentration-dependent oligomerization of cross-linked complexes between ferredoxin and ferredoxin-NADP+ reductase (2013)
  • Author(s): Yoko Kimata-Ariga, Hisako Kubota-Kawai, Young-Ho Lee, Norifumi Muraki, Takahisa Ikegami, Genji Kurisu, Toshiharu Hase
  • Journal Title: Biochemical and Biophysical Research Communications Volume: in press Issue: 4 Pages: 867-872
  • DOI: 10.1016/j.bbrc.2013.04.033
  • Peer Reviewed
• [Journal Article] Identification of a new interaction mode between the Src homology 2 domain of C-terminal Src kinase (Csk) and Csk-binding protein/phosphoprotein associated with glycosphingolipid microdomains (2013)
  • Author(s): Tanaka H, Akagi K, Oneyama C, Tanaka M, Sasaki Y, Kanou T, Lee YH, Yokogawa D, Dobenecker MW, Nakagawa A, Okada M, Ikegami T
  • Journal Title: J Biol Chem 2013 Volume: 288(21) Issue: 21 Pages: 15240-54
  • DOI: 10.1074/jbc.m112.439075
  • NAID: 120006770869
  • Peer Reviewed
• [Presentation] Toward establishing the thermodynamics of protein misfolding and aggregation (2015)
  • Author(s): Young-Ho Lee
  • Organizer: 先端融合科学シンポジウム「生体分子のダイナミクスを眺める」
  • Place of Presentation: Kobe, Japan
  • Year and Date: 2015-01-19 – 2015-01-20
• [Presentation] Finely-tuned intermolecular interactions for protein function (2015)
  • Author(s): Young-Ho Lee
  • Organizer: Korea-Japan joint international seminar
  • Place of Presentation: Osaka, Japan
  • Year and Date: 2015-01-11
• [Presentation] Finely-tuned intermolecular interactions for protein function (2014)
  • Author(s): Young-Ho Lee
  • Organizer: The 11th Japan-Korea Bilateral Symposium on Biological NMR symposium
  • Place of Presentation: Osaka, Japan
  • Year and Date: 2014-12-17 – 2014-12-20
• [Presentation] Thermodynamic characterization of supersaturation-limited protein misfolding and aggregation by calorimetry (2014)
  • Author(s): Young-Ho Lee
  • Organizer: International Seminar on "Mechanism and regulation of aberrant protein aggregation"
  • Place of Presentation: Osaka, Japan
  • Year and Date: 2014-11-17 – 2014-11-20
• [Presentation] Heat of supersaturation-limited amyloid burst directly monitored by isothermal titration calorimetry (2014)
  • Author(s): Young-Ho Lee
  • Organizer: The 14th Annual Meeting of the Protein Science Society of Japan
  • Place of Presentation: Yokohama, Japan
  • Year and Date: 2014-06-25 – 2014-06-27
• [Presentation] Cold denaturation of alpha-synuclein amyloid fibrils (2014)
  • Author(s): Young-Ho Lee(a), Tatsuya Ikenoue, and Yuji Goto(b)
  • Organizer: The 4th Conference of the Asia Pacific Protein Association (APPA2014)
  • Place of Presentation: Jeju (韓国)
• [Presentation] Intermolecular electrostatic interaction between ferredoxin and sulfite reductase studied by isothermal titration calorimetry and nuclear magnetic resonance (2014)
  • Author(s): Young-Ho Lee(a), Ju Yaen Kim, Takahisa Ikegami, Yuji Goto, and Toshiharu Hase(b)
  • Organizer: The 9th international workshop on sulfur in plants
  • Place of Presentation: Freiberg（ドイツ）
• [Presentation] Cold-denaturation of amyloid fibrils of alpha-synuclein based on a main-chain dominated architecture (2013)
  • Author(s): 李 映昊(a)
  • Organizer: 第86回日本生化学会大会
  • Place of Presentation: 横浜
  • Invited
• [Presentation] Cold-denaturation of amyloid fibrils of alpha-synuclein based on a main-chain dominated architecture (2013)
  • Author(s): Young-Ho Lee(a)
  • Organizer: Institute for Protein Research seminar on Exploring the New Horizons of Protein Folding and Misfolding Studies
  • Place of Presentation: 大阪
  • Invited
• [Book] Sulfur Metabolism in Plants (Molecular Physiology and Ecophysiology) (2015)
  • Author(s): Ju Yaen Kim, Takahisa Ikegami, Yuji Goto, Toshiharu Hase, and Young-Ho Lee
  • Total Pages: 300
  • Publisher: Springer