2014 Fiscal Year Annual Research Report
Study on effect of electrostatic interactions on stability of electron transfer complexes and amyloid fibrils
| Project/Area Number |
25870407
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| Research Institution | Osaka University |
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Principal Investigator |
李 映昊 大阪大学, たんぱく質研究所, 講師 (70589431)
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| Project Period (FY) |
2013-04-01 – 2015-03-31
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| Keywords | Amyloid fibril / Binding thermodynamics / Calorimetry / Electron transfer / Electrostatics / Enzyme activity / Interprotein interaction / Protein misfolding |
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| Outline of Annual Research Achievements |
In order to understand contribution of electrostatic interactions to formation of electron transfer complex and their effects on enzymatic activity, 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 isothermal titration calorimetry, NMR spectroscopy, and X-ray crystallographic analysis together 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. In addition, non-electrostatic interactions is also important for fine tuning of binding modes between Fd and SiR to optimize SiR activity (Kim et al. (2015) and Kim et al. in preparation). The same results were obtained in the binding system between Fd and FNR (Kinoshita et al. (2015) under peer-review).
Simultaneously, we have investigated the conformational stability of amyloid fibrils of various amyloidogenic proteins/peptides such as alpha-synuclein and amyloid beta peptides in the wide temperature range (273-385K). Although all fibrils heat-denatured over 333K, only amyloid fibrils of alpha-synuclein cold-denatured below 293K due to the repulsive electrostatic interactions at low temperature (Ikenoue and Lee et al. (2014)). Effects of maturation of alpha-synuclein fibrils on stability have been further being examined.
We have clearly demonstrated that interprotein electrostatic interactions play an important roles in normal protein functions and in clearance of disease-related amyloid fibrils.
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Research Products
(18 results)
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[Journal Article] Optimized Co-Solute Paramagnetic Relaxation Enhancement for the Rapid NMR Analysis of a Highly Fibrillogenic Peptide2015
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
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Journal Title
Journal of Biomolecular NMR
Volume: ー
Pages: 印刷中
DOI
Peer Reviewed
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[Journal Article] Ultrasonication-Dependent Formation and Degradation of Alpha-Synuclein Amyloid Fibrils2015
Author(s)
Hisashi Yagi, Aiko Mizuno, Masatomo So, Miki Hirano, Masayuki Adachi, Yoko Akazawa-Ogawa, Yoshihisa Hagihara, Tatsuya Ikenoue, Young-Ho Lee, Yasushi Kawata, and Yuji Goto
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Journal Title
Biochimica et Biophysica Acta
Volume: 1854
Pages: 209-217
DOI
Peer Reviewed
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[Journal Article] NMR Characterization of the Interaction of the Endonuclease Domain of MutL with Divalent Metal Ions and ATP2014
Author(s)
Ryota Mizushima, Ju Yaen Kim, Isao Suetake, Hiroaki Tanaka, Tomoyo Takai, Narutoshi Kamiya, Yu Takano, Yuichi Mishima, Shoji Tajima, Yuji Goto, Kenji Fukui, and Young-Ho Lee
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Journal Title
PLoS ONE
Volume: 9
Pages: e98554
DOI
Peer Reviewed / Open Access
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