Understanding the mechanism of amyloid fibrillation on the basis of single fibril and real-time observation

• Project/Area Number: 21370044
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: Osaka University
• Principal Investigator: GOTO Yuji) 大阪大学, 蛋白質研究所, 教授 (40153770)
• Co-Investigator(Kenkyū-buntansha): SAKURAI Kazumasa 大阪大学, 蛋白質研究所, 助教 (10403015)
• Co-Investigator(Renkei-kenkyūsha): FUJIWARA Toshimichi 大阪大学, 蛋白質研究所, 教授 (20242381) ; CHATANI Eri 神戸大学, 大学院・理学研究科, 准教授 (00432493) ; NAIKI Hironobu 福井大学, 医学部, 教授 (10227704)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥18,200,000 (Direct Cost: ¥14,000,000、Indirect Cost: ¥4,200,000); Fiscal Year 2011: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000); Fiscal Year 2010: ¥6,890,000 (Direct Cost: ¥5,300,000、Indirect Cost: ¥1,590,000); Fiscal Year 2009: ¥8,060,000 (Direct Cost: ¥6,200,000、Indirect Cost: ¥1,860,000)
• Keywords: 変性とフォールディング / アミロイド線維 / 蛋白質の凝集 / 蛍光顕微鏡 / 超音波 / アミロイド蛋白質 / 脳神経疾患 / 透析アミロイドーシス / アルツハイマー病 / 糖尿病 / NMR

Research Abstract

We studied the mechanism of amyloid fibrillation with a unique technique in which total internal reflection fluorescence microscopy is combined with amyloid-specific ThT. To gain insights into the possible kinetic intermediates, we performed hydrogen/ deuterium exchange of amide protons during fibril elongation. Combining the use of ultrasonication and a microplate reader, we propose an efficient approach to studying the potential of proteins to form amyloid fibrils. Taken all results together, we propose that the amyloid fibrils formed via a nucleation-dependent mechanism in a supersaturated solution, analogous to crystallization.

Research Products

• [Journal Article] Ultrasonication-dependent acceleration of amyloid fibril formation (2011)
  • Author(s): So, M., Yagi, H., Sakurai, K., Ogi, H., Naiki, H. & Goto, Y.
  • Journal Title: J. Mol. Biol. Volume: 412 Pages: 568-577
  • Peer Reviewed
• [Journal Article] Hexafluoroisopropanol induces amyloid fibrils of islet amyloid polypeptide by enhancing both hydrophobic and electrostatic interactions (2011)
  • Author(s): Yanagi, K., Ashizaki, M., Yagi, H., Sakurai, K., Lee, Y. H. & Goto, Y.
  • Journal Title: J. Biol. Chem. Volume: 286 Pages: 23959-23966
  • NAID: 120006557643
  • Peer Reviewed
• [Journal Article] Destruction of amyloid fibrils of keratoepithelin peptides by laser irradiation coupled with amyloid-specific thioflavin (2011)
  • Author(s): T. Ozawa, D., Kaji, Y., Yagi, H., Sakurai, K., Kawakami, T., Naiki, N. & Goto, Y.
  • Journal Title: J. Biol. Chem. Volume: 286 Pages: 10856-10863
  • Peer Reviewed
• [Journal Article] Seed-dependent deposition behavior of Aβpeptides studied with wireless quartz-crystal-microbalance biosensor (2011)
  • Author(s): Ogi, H., Fukunishi, Y., Yanagida, T., Yagi, H., Goto, Y., Fukushima, M., Uesugi, K., Hirao
  • Journal Title: M. Anal. Chem. Volume: 83 Pages: 4982-4988
  • NAID: 120007148353
  • Peer Reviewed
• [Journal Article] A circumventing role for the non-native intermediate in the folding ofβ-lactoglobulin (2011)
  • Author(s): Sakurai, K., Fujioka, S., Konuma, T., Yagi, M. & Goto, Y.
  • Journal Title: Biochemistry Volume: 50 Pages: 6498-6507
  • Peer Reviewed
• [Journal Article] Binding energetics of ferredoxin-NADP+ reductase with ferredoxin and its relation to function (2011)
  • Author(s): Lee, Y.-H., Ikegami, T., Standley, D. M., Sakurai, K., Hase, T. & Goto, Y.
  • Journal Title: ChemBio. Chem. Volume: 12 Pages: 2062-2070
  • Peer Reviewed
• [Journal Article] Reversible heat-induced dissociation ofβ2-microglobulin amyloid fibrils (2011)
  • Author(s): Kardos, J., Micsonai, A., Pa. l-Ga. bor, H., Petrik, E., Gra. f, L., Kova. cs, J., Lee, Y. H., Naiki, H. & Goto, Y.
  • Journal Title: Biochemistry Volume: 50 Pages: 3211-3220
  • Peer Reviewed
• [Journal Article] Inhibition of. 2-microglobulin amyloid fibril formation byα2-macroglobulin (2011)
  • Author(s): Ozawa, D., Hasegawa, K., Lee, Y. H., Sakurai, K., Yanagi, K., Ookoshi, T., Goto, Y., Naiki, H.
  • Journal Title: J. Biol. Chem. Volume: 286 Pages: 9668-9676
  • Peer Reviewed
• [Journal Article] Kinetic intermediates ofβ2-microglobulin fibril elongation probed by pulse-labeling H/ D exchange combined with NMR analysis (2011)
  • Author(s): Konuma, T., Chatani, E., Yagi, M., Sakurai, K., Ikegami, T., Naiki, H. & Goto, Y.
  • Journal Title: J. Mol. Biol. Volume: 405 Pages: 851-862
  • Peer Reviewed
• [Journal Article] Ultrasonication-dependent acceleration of amyloid fibril formation (2011)
  • Author(s): So, M., et al., Goto, Yuji
  • Journal Title: J.Mol.Biol. Volume: 412(4) Issue: 4 Pages: 568-577
  • DOI: 10.1016/j.jmb.2011.07.069
  • Peer Reviewed
• [Journal Article] Destruction of amyloid fibrils of keratoepithelin peptides by laser irradiation coupled with amyloid-specific thioflavin T. (2011)
  • Author(s): Ozawa, D., et al., Goto, Yuji
  • Journal Title: J.Biol.Chem. Volume: 286(12) Issue: 12 Pages: 10856-10863
  • DOI: 10.1074/jbc.m111.222901
  • NAID: 120006557644
  • Peer Reviewed
• [Journal Article] Hexafluoroisopropanol induces amyloid fibrils of islet amyloid polypeptide by enhancing both hydrophobic and electrostatic interactions (2011)
  • Author(s): Yanagi, K., et al., Goto, Yuji
  • Journal Title: J.Biol.Chem. Volume: 286(27) Issue: 27 Pages: 23959-23966
  • DOI: 10.1074/jbc.m111.226688
  • NAID: 120006557643
  • Peer Reviewed
• [Journal Article] Laser-induced propagation and destruction of amyloidβfibrils (2010)
  • Author(s): Yagi, H., Ozawa, D., Sakurai, K., Kawakami, T., Kuyama, H., Nishimura, O., Shimanouchi, T., Kuboi, R., Naiki, H. & Goto, Y.
  • Journal Title: J. Biol. Chem. Volume: 285 Pages: 19660-19667
  • NAID: 120006557646
  • Peer Reviewed
• [Journal Article] Amyloid fibrils of the constant domain of immunoglobulin light chain (2010)
  • Author(s): Yamamoto, K., Yagi, H., Lee, Y. H., Kardos, K., Hagihara, Y., Naiki, H. & Goto, Y.
  • Journal Title: FEBS Lett. Volume: 584 Pages: 3348-3353
  • Peer Reviewed
• [Journal Article] Isolation of short peptide fragments fromα-synuclein fibril core identifies a residue important for fibril nucleation : a possible implication for diagnostic applications (2010)
  • Author(s): Yagi, H., Takeuchi, H., Ogawa, S., Ito, N., Sakane, I., Hongo, K., Mizobata, T., Goto, Y. & Kawata, Y.
  • Journal Title: Biochim. Biophys. Acta Volume: 1804 Pages: 2077-2087
  • Peer Reviewed
• [Journal Article] アミロイドーシス発症の分子機構解明 (2010)
  • Author(s): 八木寿梓、後藤祐児
  • Journal Title: メディカルバイオ Volume: 7 Pages: 30-31
  • Peer Reviewed
• [Journal Article] Direct observation of minimum-sized amyloid fibrils using solution NMR spectroscopy (2010)
  • Author(s): Yoshimura, Y., Sakurai, K., Lee, Y.-H., Ikegami, T., Chatani, E., Naiki, H. & Goto, Y.
  • Journal Title: Protein Sci. Volume: 19 Pages: 2347-2355
  • Peer Reviewed
• [Journal Article] Pre-steady state kinetic analysis for the elongation of amyloid fibrils ofβ2-microglobulin with tryptophan mutagenesis (2010)
  • Author(s): Chatani, E., Ohnishi1, R., Konuma, T., Sakurai, K., Naiki, N. & Goto, Y.
  • Journal Title: J. Mol. Biol. Volume: 400 Pages: 1057-1066
  • Peer Reviewed
• [Journal Article] タンパク質のアミロイド形成-過飽和条件で揺らぎが引きおこす晶析現象 (2010)
  • Author(s): 後藤祐児、李映昊
  • Journal Title: メディカルバイオ10月号別冊(寺嶋正秀監修) Pages: 38-34
  • Peer Reviewed
• [Journal Article] Laser-induced propagation and destruction of amyloid β fibrils. (2010)
  • Author(s): Yagi, Hisashi ; Goto, Yuji
  • Journal Title: J.Biol.Chem. Volume: 285(25) Pages: 19660-19667
  • NAID: 120006557646
  • Peer Reviewed
• [Journal Article] Pre-steady state kinetic analysis for the elongation of amyloid fibrils of β2-microglobulin with tryptophan mutagenesis. (2010)
  • Author(s): Chatani, Eri ; Goto, Yuji
  • Journal Title: J.Mol.Biol. Volume: 400(5) Pages: 1257-1066
  • Peer Reviewed
• [Journal Article] Direct observation of minimum-sized amyloid fibrils using solution NMR spectroscopy. (2010)
  • Author(s): Yoshimura, Yuichi ; Goto, Yuji
  • Journal Title: Protein Sci. Volume: 19(12) Pages: 2347-2355
  • Peer Reviewed
• [Journal Article] Critical role of interfaces and agitation on the nucleation of Aβ amyloid fibrils at low concentrations of Aβ monomers (2010)
  • Author(s): Morinaga, A., et al.
  • Journal Title: Biochim.Biophys.Acta 1804(4) Pages: 986-995
  • Peer Reviewed
• [Journal Article] Differences in the molecular structure of β2-microglobulin between two morphologically different amyloid fibrils (2010)
  • Author(s): Hiramatsu, H., et al.
  • Journal Title: Biochemistry 49(4) Pages: 742-751
  • Peer Reviewed
• [Journal Article] Ultrasonication establishes the equilibrium between production and breakdown leading to the minimum-sized amyloid fibrils (2009)
  • Author(s): Chatani, E., Lee, Y.-H., Yagi, H., Yoshimura, Y., Naiki, N. & Goto, Y.
  • Journal Title: Proc. Natl. Acad. Sci. USA Volume: 106 Pages: 11119-11124
  • Peer Reviewed
• [Journal Article] Thermal response with exothermic effects of. 2-microglobulin amyloid fibrils and fibrillation (2009)
  • Author(s): Sasahara, K., Yagi, H., Naiki, H. & Goto, Y.
  • Journal Title: J. Mol. Biol. Volume: 389 Pages: 584-594
  • Peer Reviewed
• [Journal Article] Destruction of amyloid fibrils of aβ2-microglobulin fragment by laser beam irradiation (2009)
  • Author(s): Ozawa, D., Yagi, H., Ban, T., Kameda, A., Kawakami, T., Naiki, H., & Goto, Y.
  • Journal Title: J. Biol. Chem. Volume: 284 Pages: 1009-1017
  • Peer Reviewed
• [Journal Article] Branching in amyloid fibril growth. Andersen (2009)
  • Author(s): C. B., Yagi, H., Manno, M., Martorana, V., Ban, T., Christiansen, G., Otzen, D., Goto, Y. & Rischel, C.
  • Journal Title: Biophys. J. Volume: 96 Pages: 1529-1536
  • Peer Reviewed
• [Journal Article] NMR-based characterization of a refolding intermediate ofβ2-microglobulin labeled amino acid selectively using a wheat germ cell-free system (2009)
  • Author(s): Kameda, A., Morita, E.-H., Sakurai, K., Naiki, N. & Goto, Y.
  • Journal Title: Protein Sci. Volume: 18 Pages: 1592-1601
  • Peer Reviewed
• [Journal Article] Structural dynamics and folding ofβ-lactoglobulin probed by heteronuclear NMR (2009)
  • Author(s): Sakurai, K., Konuma, T., Yagi, M. & Goto, Y.
  • Journal Title: Biochim. Biophys. Acta Volume: 1790 Pages: 527-537
  • Peer Reviewed
• [Journal Article] A comprehensive model for packing and hydration for amyloid fibrils of. 2-microglobulin (2009)
  • Author(s): Lee, Y.-H., Chatani, E., Sasahara, K., Naiki, H. & Goto, Y.
  • Journal Title: J. Biol. Chem. Volume: 284 Pages: 2169-2175
  • Peer Reviewed
• [Journal Article] Thermal response with exothermic effects of β2-microglobulin amyloid fibrils and fibrillation. (2009)
  • Author(s): Sasahara, K., et al.
  • Journal Title: J.Mol.Biol. 389(3) Pages: 584-594
  • Peer Reviewed
• [Journal Article] Thermal response with exothermic effects of β2-microglobulin amyloid fibrils and fibrillation. (2009)
  • Author(s): Pal-Gabor, H., et al.
  • Journal Title: Biochemistry 48(24), Pages: 5689-5699
  • Peer Reviewed
• [Journal Article] NMR-based characterization of a refolding intermediate of β2-microglobulin labeled amino acid selectively using a wheat germ cell-free system. (2009)
  • Author(s): Kameda, A., et al.
  • Journal Title: Protein Science 18(8) Pages: 1592-1601
  • Peer Reviewed
• [Journal Article] Ultrasonication establishes the equilibrium between production and breakdown leading to the minimum-sized amyloid fibrils (2009)
  • Author(s): Chatani, E., et al.
  • Journal Title: Proc.Natl.Acad.Sci.USA 106(27) Pages: 11119-11124
  • Peer Reviewed
• [Presentation] 「蛋白質のフォールディングと異常凝集の一般原理の解明を目指して」 (2011)
  • Author(s): 後藤祐児
  • Organizer: 関西学院大学・理工学部50周年記念講演会、文科省オープン・リサーチセンター成果報告会「ナノマテリアルと生体分子の物理化学」
  • Place of Presentation: 関西学院大学(西宮)
  • Year and Date: 2011-10-08
• [Presentation] 蛋白質のフォールディングと異常凝集の一般原理の解明を目指して (2011)
  • Author(s): 後藤祐児
  • Organizer: 関西学院大学・理工学部50周年記念講演会、文科省オープン・リサーチセンター成果報告会「ナノマテリアルと生体分子の物理化学」
  • Place of Presentation: 関西学院大学、西宮(招待講演)
  • Year and Date: 2011-10-08
• [Presentation] Toward understanding a unifying principle of protein folding and aggregation (2011)
  • Author(s): Goto, Y.
  • Organizer: The First Korean Protein Society Symposium
  • Place of Presentation: Seoul National University, Seoul, Korea
  • Year and Date: 2011-09-23
• [Presentation] Toward understanding a unifying principle of protein folding and aggregation (2011)
  • Author(s): 後藤祐児
  • Organizer: The First Korean Protein Society Symposium
  • Place of Presentation: Seoul National University, Seoul, Korea(招待講演)
  • Year and Date: 2011-09-23
• [Presentation] 「蛋白質のフォールディングとアミロイド線維形成」 (2011)
  • Author(s): 後藤祐児
  • Organizer: 日本物理学会2011年周期大会シンポジウム「巨大分子～サブミクロン粒子の自己集積」
  • Place of Presentation: 富山大学
  • Year and Date: 2011-09-21
• [Presentation] 蛋白質のフォールディングとアミロイド線維形成 (2011)
  • Author(s): 後藤祐児
  • Organizer: 日本物理学会2011年周期大会シンポジウム「巨大分子～サブミクロン粒子の自己集積」
  • Place of Presentation: 富山大学、富山(招待講演)
  • Year and Date: 2011-09-21
• [Presentation] Toward understanding a unifying principle of protein folding and aggregation (2011)
  • Author(s): Goto, Y.
  • Organizer: International Symposium of the Protein Society of Thailand
  • Place of Presentation: Chulabhorn Conference Center, Bangkok, Thailand
  • Year and Date: 2011-08-31
• [Presentation] Toward understanding a unifying principle of protein folding and aggregation (2011)
  • Author(s): 後藤祐児
  • Organizer: International Symposium of the Protein Society of Thailand
  • Place of Presentation: Chulabhorn Conference Center, Bangkok, Thailand(招待講演)
  • Year and Date: 2011-08-31
• [Presentation] Visualization of amyloid fibril formation of proteins. (2010)
  • Author(s): Goto, Yuji
  • Organizer: 10^<th> KIAS Conference on Protein Structure and Function
  • Place of Presentation: KIAS, Seoul, Korea
  • Year and Date: 2010-10-02
• [Presentation] Amyloid fibril growth visualized by thioflavin T fluorescence and probed by H/ D exchange (2010)
  • Author(s): Goto, Y.
  • Organizer: Gordon Research Conference : Protein Folding Dynamics
  • Place of Presentation: Ventura CA, USA
• [Presentation] Amyloid fibril growth visualized by thioflavin T fluorescence and probed by H/ D exchange (2010)
  • Author(s): Goto, Y.
  • Organizer: Symposium on Structure and Folding of Disease Related Proteins
  • Place of Presentation: Seoul National University, Seoul, Korea
• [Presentation] Visualization of amyloid fibril formation of proteins (2010)
  • Author(s): Goto, Y.
  • Organizer: 10^<th> KIAS Conference on Protein Structure and Function
  • Place of Presentation: Seoul, Korea
• [Presentation] Amyloid fibril growth visualized by thioflavin T fluorescence and probed by H/D exchange. (2010)
  • Author(s): Goto, Yuji
  • Organizer: Gordon Research Conference : Protein Folding Dynamics,
  • Place of Presentation: Ventura, USA
• [Presentation] Amyloid fibril growth visualized by thioflavin T fluorescence and probed by H/D exchange. (2009)
  • Author(s): Goto, Yuji
  • Organizer: The 23rd Symposium of The Protein Society
  • Place of Presentation: Boston, USA
  • Year and Date: 2009-07-26
• [Presentation] Folding and amyloid fibril formation of proteins. (2009)
  • Author(s): Goto, Yuji
  • Organizer: The 66th KSBMB Annual Meeting
  • Place of Presentation: Seoul, Korea
  • Year and Date: 2009-05-12
• [Presentation] Amyloid fibril growth visualized by thioflavin T fluorescence and probed by H/ D exchange (2009)
  • Author(s): Goto, Y.
  • Organizer: International Bunsen Discussion Meeting on Structure of Amyloid Fibrils and Mechanism of Amyloid Formation
  • Place of Presentation: Universitat Halle, Halle an der Saale, Germany
• [Presentation] Amyloid fibril growth visualized by thioflavin T fluorescence and probed by H/ D exchange (2009)
  • Author(s): Goto, Y.
  • Organizer: The 23rd Symposium of The Protein Society
  • Place of Presentation: Boston, USA
• [Presentation] Amyloid fibril growth visualized by thioflavin T fluorescence and probed by H/ D exchange (2009)
  • Author(s): Goto, Y.
  • Organizer: The 3rd Asian-Pacific Peptide Symposium
  • Place of Presentation: Jeju island, Korea
• [Book] Real-time observation of amyloid-βfibril growth by total internal reflection fluorescence miscroscopy (2010)
  • Author(s): Ban, T. & Goto, Y.
  • Publisher: Protein Misfolding Disease
• [Book] Water and Biomolecules : Physical Chemistry of Life Phenomena (eds. Kuwajima K, Goto Y, Hirata F, Kataoka M, & Terazima M) (2009)
  • Author(s): Yagi, H., et al.(分担執筆)
  • Publisher: Springer-Verlag, Berlin and Heidelberg, Germany
• [Remarks]
  • URL: http://www.protein.osaka-u.ac.jp/physical/yoeki.html
• [Remarks]
  • URL: http://www.protein.osaka-u.ac.jp/physical/yoeki.html
• [Remarks]
  • URL: http://www.protein.osaka-u.ac.jp/physical/yoeki.html