Development of analytical techniques for structural property dynamics based on high-speed AFM

• Project Area: Soft Crystals: Science and Photofunctions of Easy-Responsive Systems with Felxibility and Higher-Ordering
• Project/Area Number: 18H04512
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Research Institution: Nagoya University
• Principal Investigator: 内橋 貴之 名古屋大学, 理学研究科, 教授 (30326300)
• Project Period (FY): 2018-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2019: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000); Fiscal Year 2018: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
• Keywords: 高速原子間力顕微鏡 / 力学マッピング / ゲル微粒子 / ヘムタンパク質 / 結晶構造 / 機械特性 / ダイナミクス解析 / 超分子ファイバー / ソフトクリスタル / ナノ動態 / 構造 / ダイナミクス評価

Outline of Annual Research Achievements

本研究課題では、高速AFMを用いて外部刺激に応答するソフトクリスタルの表面・物性構造のダイナミクスを高時空間分解能で可視化する基盤技術を確立し、ソフトクリスタルの機能創発の分子機構を明らかにすることを目的としている。以下に今年度の成果を項目ごとに記述する。
1. 高速力学マッピング法の開発：高速AFMでイメージング中に任意の位置での定量機械特性を取得可能な”in-line-force-curve”法を開発した。この手法を用いてチューブリンが重合したタンパク質ポリマーである微小管の単一チューブリンダイマーの引き抜きによる局所構造操作に成功し、チューブリンダイマー間の結合エネルギーの定量化を行った。さらに、”in-line-force-curve”を拡張し、機械特性(弾性率・凝着力)を1フレーム3秒程度でマッピングできるシステムを開発した。
2. 高分子ゲル微粒子の昇温収縮の過程：温度制御型高速AFMと位相計測を組み合わせて、熱応答性のpNiPAmハイドロゲル微粒子の温度依存的収縮過程を観察したところ、微粒子内部に非熱応答性のナノサイズのドメインが存在することを見出した。また、この非熱応答性ドメインは沈殿重合法、あるいは体積相転移温度以下での逆相ミニエマルション重合法で重合した微粒子に存在し、体積相転移温度よりも高い温度で重合された微粒子ではナノドメイン微粒子内に均一に分布することを明らかにした。
3. 六量体ヘムタンパク質のシート構造形成過程の観察：六量体ヘムタンパク質(HTTP)のモノマーにCys残基を導入してトリペプチドFGGタグを付与し、さらに、ククルビットウリル8(CB8)をFGGタグのホストとすることで、HTTPのシート状自己組織化構造を作製することに成功した。また、シート形成過程を高速AFMでモニターし、シート構造形成の詳細を明らかにした。

Research Progress Status

令和元年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

令和元年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Recent advances in bioimaging with high-speed atomic force microscopy (2020)
  • Author(s): Uchihashi Takayuki、Ganser Christian
  • Journal Title: Biophysical Reviews Volume: 12 Issue: 2 Pages: 363-369
  • DOI: 10.1007/s12551-020-00670-z
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Dynamics of oligomer and amyloid fibril formation by yeast prion Sup35 observed by high-speed atomic force microscopy (2020)
  • Author(s): Konno H, Watanabe-Nakayama T, Uchihashi T, Okuda M, Zhu L, Kodera N, Kikuchi Y, Ando T, Taguchi H
  • Journal Title: Proc. Natl. Acad. Sci. U S A Volume: 117 Issue: 14 Pages: 7831-7836
  • DOI: 10.1073/pnas.1916452117
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Construction of a Hexameric Hemoprotein Sheet and Direct Observation of Dynamic Processes of Its Formation (2020)
  • Author(s): Oohora Koji、Hirayama Shota、Uchihashi Takayuki、Hayashi Takashi
  • Journal Title: Chemistry Letters Volume: 49 Issue: 2 Pages: 186-190
  • DOI: 10.1246/cl.190855
  • NAID: 130007795242
  • Peer Reviewed
• [Journal Article] Rad50 zinc hook functions as a constitutive dimerization module interchangeable with SMC hinge (2020)
  • Author(s): Tatebe Hisashi、Lim Chew Theng、Konno Hiroki、Shiozaki Kazuhiro、Shinohara Akira、Uchihashi Takayuki、Furukohri Asako
  • Journal Title: Nature Communications Volume: 11 Issue: 1 Pages: 370-370
  • DOI: 10.1038/s41467-019-14025-0
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Thermoresponsive Micellar Assembly Constructed from a Hexameric Hemoprotein Modified with Poly(N-isopropylacrylamide) toward an Artificial Light-Harvesting System (2020)
  • Author(s): Hirayama Shota、Oohora Koji、Uchihashi Takayuki、Hayashi Takashi
  • Journal Title: Journal of the American Chemical Society Volume: 142 Issue: 4 Pages: 1822-1831
  • DOI: 10.1021/jacs.9b10080
  • Peer Reviewed / Open Access
• [Journal Article] Supramolecular tholos-like architecture constituted by archaeal proteins without functional annotation. (2020)
  • Author(s): Maho Yagi-Utsumi,Arunima Sikdar,Chihong Song,Jimin Park,Rintaro Inoue,Hiroki Watanabe,Raymond N Burton-Smith,Toshiya Kozai,Tatsuya Suzuki,Atsuji Kodama,Kentaro Ishii,Hirokazu Yagi,Tadashi Satoh,Susumu Uchiyama,Takayuki Uchihashi,Keehyoung Joo,Jooyoung Lee,Masaaki Sugiyama,Kazuyoshi Murata,Koichi Kato
  • Journal Title: Scientific Reports Volume: 10 Issue: 1 Pages: 1540-1540
  • DOI: 10.1038/s41598-020-58371-2
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Non-Thermoresponsive Decanano‐sized Domains in Thermoresponsive Hydrogel Microspheres Revealed by Temperature‐Controlled High‐Speed Atomic Force Microscopy (2019)
  • Author(s): Yuichiro Nishizawa,Shusuke Matsui,Kenji Urayama,Takuma Kureha,Mitsuhiro Shibayama,Takayuki Uchihashi,Daisuke Suzuki
  • Journal Title: Angewandte Chemie International ed. Volume: 131 Issue: 26 Pages: 8901-8905
  • DOI: 10.1002/ange.201903483
  • Peer Reviewed
• [Journal Article] Hydrogel Microellipsoids that Form Robust String-like Assemblies at the Air/Water Interface (2019)
  • Author(s): Honda Kenshiro、Sazuka Yuka、Iizuka Kojiro、Matsui Shusuke、Uchihashi Takayuki、Kureha Takuma、Shibayama Mitsuhiro、Watanabe Takumi、Suzuki Daisuke
  • Journal Title: Angewandte Chemie International Edition Volume: 印刷中 Issue: 22 Pages: 7294-7298
  • DOI: 10.1002/anie.201901611
  • Peer Reviewed
• [Journal Article] Mutational and Combinatorial Control of Self-Assembling and Disassembling of Human Proteasome α Subunits (2019)
  • Author(s): Taichiro Sekiguchi, Tadashi Satoh, Eiji Kurimoto, Chihong Song, Toshiya Kozai, Hiroki Watanabe, Kentaro Ishii, Hirokazu Yagi, Saeko Yanaka, Susumu Uchiyama, Takayuki Uchihashi, Kazuyoshi Murata and Koichi Kato
  • Journal Title: Int. J. Mol. Sci. Volume: 20 Issue: 9 Pages: 2308-2308
  • DOI: 10.3390/ijms20092308
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Construction of a quadrangular tetramer and a cage-like hexamer from three-helix bundle-linked fusion proteins (2019)
  • Author(s): Miyamoto Takaaki、Hayashi Yugo、Yoshida Keito、Watanabe Hiroki、Uchihashi Takayuki、Yonezawa Kento、Shimizu Nobutaka、Kamikubo Hironari、Hirota Shun
  • Journal Title: ACS Synthetic Biology Volume: 印刷中 Issue: 5 Pages: 1112-1120
  • DOI: 10.1021/acssynbio.9b00019
  • NAID: 120006801961
  • Peer Reviewed / Open Access
• [Journal Article] The Fab portion of immunoglobulin G contributes to its binding to Fcγ receptor III. (2019)
  • Author(s): [Rina Yogo,Yuki Yamaguchi,Hiroki Watanabe,Hirokazu Yagi,Tadashi Satoh,Mahito Nakanishi,Masayoshi Onitsuka,Takeshi Omasa,Mari Shimada,Takahiro Maruno,Tetsuo Torisu,Shio Watanabe,Daisuke Higo,Takayuki Uchihashi,Saeko Yanaka,Susumu Uchiyama,Koichi Kato]
  • Journal Title: Scientific Reports Volume: 9 Issue: 1 Pages: 11957-11957
  • DOI: 10.1038/s41598-019-48323-w
  • NAID: 120006875664
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Protein uptake into individual hydrogel microspheres visualized by high-speed atomic force microscopy. (2019)
  • Author(s): Shusuke Matsui,Kensuke Hosho,Haruka Minato,Takayuki Uchihashi,Daisuke Suzuki
  • Journal Title: Chemical Communications Volume: 55 Issue: 68 Pages: 10064-10067
  • DOI: 10.1039/c9cc05116c
  • Peer Reviewed
• [Journal Article] Crystal structure of heliorhodopsin (2019)
  • Author(s): W. Shihoya, K. Inoue, M. Singh, M. Konno, S. Hososhima, K. Yamashita, K. Ikeda, A. Higuchi, T. Izume, Okazaki S, Hashimoto M, Mizutori R, Tomida S, Yamauchi Y, Abe-Yoshizumi R, K. Katayama, SP. Tsunoda, M. Shibata, Y. Furutani, A. Pushkarev, O. Béjà, T. Uchihashi, H. Kandori, O. Nureki
  • Journal Title: Nature Volume: 574 Issue: 7776 Pages: 132-136
  • DOI: 10.1038/s41586-019-1604-6
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] On-Membrane Dynamic Interplay between Anti-GM1 IgG Antibodies and Complement Component C1q. (2019)
  • Author(s): Saeko Yanaka,Rina Yogo,Hiroki Watanabe,Yuki Taniguchi,Tadashi Satoh,Naoko Komura,Hiromune Ando,Hirokazu Yagi,Nobuhiro Yuki,Takayuki Uchihashi,Koichi Kato
  • Journal Title: International Journal of Molecular Sciences Volume: 21 Issue: 1 Pages: 147-147
  • DOI: 10.3390/ijms21010147
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Structural basis of nucleosome assembly by the Abo1 AAA+?ATPase histone chaperone. (2019)
  • Author(s): Carol Cho,Juwon Jang,Yujin Kang,Hiroki Watanabe,Takayuki Uchihashi,Seung Joong Kim,Koichi Kato,Ja Yil Lee,Ji-Joon Song
  • Journal Title: Nature Communications Volume: 10 Issue: 1 Pages: 5764-5764
  • DOI: 10.1038/s41467-019-13743-9
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Microtubule self-healing and defect creation investigated by in-line force measurements during high-speed atomic force microscopy imaging (2019)
  • Author(s): Ganser Christian、Uchihashi Takayuki
  • Journal Title: Nanoscale Volume: 11 Issue: 1 Pages: 125-135
  • DOI: 10.1039/c8nr07392a
  • Peer Reviewed
• [Journal Article] A ring-shaped hemoprotein trimer thermodynamically controlled by the supramolecular heme-heme pocket interaction (2019)
  • Author(s): Oohora Koji、Kajihara Ryota、Fujimaki Nishiki、Uchihashi Takayuki、Hayashi Takashi
  • Journal Title: Chemical Communications Volume: 55 Issue: 11 Pages: 1544-1547
  • DOI: 10.1039/c8cc09314h
  • Peer Reviewed
• [Journal Article] Supramolecular Hemoprotein Assembly with a Periodic Structure Showing Heme-Heme Exciton Coupling (2018)
  • Author(s): Oohora Koji、Fujimaki Nishiki、Kajihara Ryota、Watanabe Hiroki、Uchihashi Takayuki、Hayashi Takashi
  • Journal Title: Journal of the American Chemical Society Volume: 140 Issue: 32 Pages: 10145-10148
  • DOI: 10.1021/jacs.8b06690
  • Peer Reviewed
• [Journal Article] Monitoring Thermoresponsive Morphological Changes in Individual Hydrogel Microspheres (2018)
  • Author(s): Matsui Shusuke、Nishizawa Yuichiro、Uchihashi Takayuki、Suzuki Daisuke
  • Journal Title: ACS Omega Volume: 3 Issue: 9 Pages: 10836-10842
  • DOI: 10.1021/acsomega.8b01770
  • Peer Reviewed / Open Access
• [Journal Article] Direct Observation and Manipulation of Supramolecular Polymerization by High-Speed Atomic Force Microscopy (2018)
  • Author(s): Fukui Tomoya、Uchihashi Takayuki、Sasaki Norihiko、Watanabe Hiroki、Takeuchi Masayuki、Sugiyasu Kazunori
  • Journal Title: Angewandte Chemie International Edition Volume: 57 Issue: 47 Pages: 15465-15470
  • DOI: 10.1002/anie.201809165
  • Peer Reviewed
• [Presentation] Imaging of Single-Molecule Dynamics using High-Speed Atomic Force Microscopy (2019)
  • Author(s): Takayuki Uchihashi
  • Organizer: The 2nd NIBB-Princeton Joint Symposium on Imaging and Quantitative Biology
  • Int'l Joint Research / Invited
• [Presentation] Real-Time Visualization of Biological Molecules at Work with High-Speed Atomic Force Microscopy (2019)
  • Author(s): Takayuki Uchihashi
  • Organizer: IEEE International Conference on Manipulation, Manufacturing and Measurement on the Nanoscale 2019
  • Int'l Joint Research / Invited
• [Presentation] Nanoscale Dynamic Imaging of Biological Molecules with High-Speed Atomic Force Microscopy (2019)
  • Author(s): Takayuki Uchihashi
  • Organizer: Frontier Bioorganization Forum 2019
  • Int'l Joint Research / Invited
• [Presentation] High-Speed Atomic Force Microscopy for Direct Visualization from Single Protein to Cell Dynamics (2019)
  • Author(s): Takayuki Uchihashi
  • Organizer: EsCELLS visit talks
  • Int'l Joint Research / Invited
• [Presentation] Defect creation in microtubules and kinesin motility studied by HS-AFM (2019)
  • Author(s): Christian Ganser, Kimitoshi Takeda, Ryota Iino, Koichi Kato and Takayuki Uchihashi
  • Organizer: 27th International Colloquium on Scanning Probe Microscopy (ICSPM27)
  • Int'l Joint Research
• [Presentation] Real-Time Monitoring of Morphological Change of Metal Surface During Corrosion Process with High-Speed Atomic Force Microscopy (2019)
  • Author(s): Takayuki Uchihashi, Mitsuhiro Okada, Miyuki Tanaka, Toshitaka Kubo, Tetsuo Shimizu
  • Organizer: 27th International Colloquium on Scanning Probe Microscopy (ICSPM27)
  • Int'l Joint Research
• [Presentation] Kinesin transport on microtubules studied by HS-AFM (2019)
  • Author(s): Christian Ganser and Takayuki Uchihashi
  • Organizer: The 2nd NIBB-Princeton Joint Symposium on Imaging and Quantitative Biology
  • Int'l Joint Research
• [Presentation] Dynamics of photosystem II protein complexes as observed by high-speed atomic force microscopy (2019)
  • Author(s): Takaya Tokano, Yuki Kato, Shogo Sugiyama, Takumi Noguchi, Takayuki Uchihashi
  • Organizer: The 2nd NIBB-Princeton Joint Symposium on Imaging and Quantitative Biology
  • Int'l Joint Research
• [Presentation] 高速原子間力顕微鏡と生体・人工分子のダイナミクス計測 (2019)
  • Author(s): 内橋貴之
  • Organizer: 第23回コロイド・界面技術者フォーラム～コロイド領域のイメージング・評価技術～
  • Invited
• [Presentation] ダイナミンの動的分子イメージング：in vitro 再構成系×高速 AFM (2019)
  • Author(s): 竹居孝二, 内橋貴之
  • Organizer: ワークショップ「新しいコラボの在り方を探る～細胞生物学ｘ蛋白質科学」 第19回 日本蛋白質科学会年会, 第71回 日本細胞生物学会大会 合同年次大会
  • Invited
• [Presentation] 溶液中高速原子間力顕微鏡による生体・人工分子のダイナミクス解析 (2019)
  • Author(s): 内橋貴之
  • Organizer: シンポジウム「原子間力顕微鏡技術のパラダイムシフト～原子から細胞まで階層をまたぐ構造・機能解析～」 日本顕微鏡学会 第75回学術講演会
• [Presentation] 高速原子間力顕微鏡による生細胞イメージング (2019)
  • Author(s): 内橋貴之
  • Organizer: 日本学術振興会 ナノプローブテクノロジー第167委員会 第91回研究会
  • Invited
• [Presentation] Dynamic Structural Analysis of Proteins at Work with High-Speed Atomic Force Microscopy (2019)
  • Author(s): 内橋貴之
  • Organizer: 【応用物理学界 関西支部】支部セミナー「バイオフォトニクス研究の最前線」
  • Invited
• [Presentation] Visualization of Single Molecule Dynamics at Work with High-Speed Atomic Force Microscopy (2019)
  • Author(s): 内橋貴之
  • Organizer: 1st ITbM Frontier Seminar
  • Invited
• [Presentation] 高速原子間力顕微鏡で可視化する分子ダイナミクス (2019)
  • Author(s): 内橋貴之
  • Organizer: 日本学術振興会 情報科学用有機材料第142委員会 合同研究会
  • Invited
• [Presentation] n-situ observation of self-assembling fiber formation of biological and artificial molecules (2018)
  • Author(s): Takayuki Uchihashi
  • Organizer: ACSIN-14 & ICSPM26
  • Int'l Joint Research
• [Presentation] Direct observation of self-assembly process of biological and artificial fibrils using high-speed atomic force microscopy (2018)
  • Author(s): Takayuki Uchihashi
  • Organizer: Interhierarchical understanding of materials and life through molecular observation
  • Int'l Joint Research / Invited
• [Presentation] High-Speed Atomic Force Microscopy for Visualization of Dynamic Processes in Biological and Artificial Supramolecules (2018)
  • Author(s): Takayuki Uchihashi
  • Organizer: International Scanning Probe Microscopy 2018
  • Int'l Joint Research / Invited
• [Presentation] High-speed Atomic Force Microscopy: A tool for direct visualization of single-molecule dynamics (2018)
  • Author(s): Takayuki Uchihashi
  • Organizer: Minisymposium on “Advanced Atomic Force Microscopy”
  • Int'l Joint Research / Invited
• [Presentation] Imaging and Manipulation of Biological Molecules with High-Speed Atomic Force Microscopy (2018)
  • Author(s): Takayuki Uchihashi
  • Organizer: ICN-T 2018
  • Int'l Joint Research / Invited
• [Presentation] High-Speed Atomic Force Microscopy for Visualization and Manipulation of Biological and Artificial Molecules (2018)
  • Author(s): Takayuki Uchihashi
  • Organizer: SPMonSPM 2018
  • Int'l Joint Research / Invited
• [Presentation] High-speed atomic force microscopy for direct visualization of biological macromolecules at work (2018)
  • Author(s): Takayuki Uchihashi
  • Organizer: The 79th Okazaki Conference "Synthetic, Biological, and Hybrid Molecular Engines"
  • Int'l Joint Research / Invited
• [Presentation] Connection between AFM data and computational simulation - image processing for quantitative analysis and demands from the experimental side - (2018)
  • Author(s): Takayuki Uchihashi
  • Organizer: Trends in Computational Molecular Biophysics Workshop
  • Int'l Joint Research / Invited
• [Presentation] 高速原子間力顕微鏡で可視化する生体分子のナノ動態 (2018)
  • Author(s): 内橋貴之
  • Organizer: NSIセミナー・アドバンス生命理学特論,
  • Invited
• [Presentation] 高速原子間力顕微鏡を用いた溶液環境下での分子のダイナミクス計測 (2018)
  • Author(s): 内橋貴之
  • Organizer: 平成30年度実践セミナー 『光・ナノ計測実践セミナーⅢ』
  • Invited
• [Presentation] 原子間力顕微鏡でリアルタイム可視化する生体/人工超分子の重合ダイナミクス (2018)
  • Author(s): 内橋貴之
  • Organizer: 日本物理学会 2018年秋季大会 領域9, 5合同シンポジウム「時間分解プローブを駆使した表面・界面科学及び結晶成長の進展と展望 」
  • Invited
• [Book] 自己修復材料、 自己組織化、形状記憶材料の開発と応用事例 (2020)
  • Author(s): 内橋貴之
  • Total Pages: 446
  • Publisher: （株）技術情報協会
  • ISBN: 9784861047817
• [Book] Compendium of Surface and Interface Analysis (2018)
  • Author(s): Surface Science Society of Japan
  • Total Pages: 853
  • Publisher: Springer
  • ISBN: 9789811061561
• [Book] イメージングの選び方・使い方100＋ (2018)
  • Author(s): 原田 慶恵、永井 健治
  • Total Pages: 236
  • Publisher: 羊土社
  • ISBN: 9784758103756
• [Remarks] 名古屋大学D研ホームページ
  • URL: http://d.phys.nagoya-u.ac.jp/index.html