T4ファージ由来蛋白質針による生体膜透過の動的秩序機構

• Project Area: Dynamical ordering of biomolecular systems for creation of integrated functions
• Project/Area Number: 16H00755
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Science and Engineering
• Research Institution: Tokyo Institute of Technology
• Principal Investigator: 上野 隆史 東京工業大学, 生命理工学院, 教授 (70332179)
• Project Period (FY): 2016-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥6,370,000 (Direct Cost: ¥4,900,000、Indirect Cost: ¥1,470,000); Fiscal Year 2017: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000); Fiscal Year 2016: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000)
• Keywords: バクテリオファージT4 / タンパク質集合体 / 超分子化学 / タンパク質工学 / 分子機械 / ファージ / タンパク質 / 生体材料 / ナノバイオ / バイオテクノロジー

Outline of Annual Research Achievements

近年、多くのファージや、分泌系では、効率的な膜輸送反応に針状蛋白質を利用していることが明らかとなり、その応用利用が注目されている。そこで、本研究では、バクテリオファージT4の部品蛋白質から合成した蛋白質針の自発的な細胞膜透過能に着目し、そのメカニズムの解明を目指した。具体的には、H20-H22年度基盤研究(B)「ウイルス部品蛋白質からなるナノチューブ構造体の分子設計と機能化」と、H26-H27年度新学術領域研究「動的秩序」「人工分子針の細胞膜貫通制御」により見出した蛋白質針の細胞膜透過現象をもとに、以下に示す三種類の実験を組み合わせた領域内外の融合研究により、蛋白質針の膜透過機能の解明を達成した。(1)高速AFM観察：研究の要となる高速AFM測定では、マイカ基板上へ吸着した人工蛋白質針を脂質二分子膜の端へ相互作用させることによって、膜貫通プロセスを二次元的な運動として1分子観察した。(2) MD計算：高速AFM解析によって得られた蛋白質針と脂質膜との接触角、接触保持時間、挿入速度をパラメーター化し、それらの構造情報をもとに、MD計算を進めた。(3)電気生理学測定：液滴接触法による脂質二分子膜作成から、迅速測定系を構築し、脂質膜透過と膜間電位差の関係を明らかとした。さらに、針先構造や表面電荷の異なる蛋白質針の合成を行い、上記の実験による膜透過能の比較から、膜透過メカニズムの全容を解明した。

Research Progress Status

29年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

29年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Engineering of protein assemblies within cells (2018)
  • Author(s): Nguyen Tien K、Ueno Takafumi
  • Journal Title: Current Opinion in Structural Biology Volume: 51 Pages: 1-8
  • DOI: 10.1016/j.sbi.2017.12.005
  • Peer Reviewed
• [Journal Article] Functionalization of protein crystals with metal ions, complexes and nanoparticles (2018)
  • Author(s): Abe Satoshi、Maity Basudev、Ueno Takafumi
  • Journal Title: Current Opinion in Chemical Biology Volume: 43 Pages: 68-76
  • DOI: 10.1016/j.cbpa.2017.11.015
  • Peer Reviewed
• [Journal Article] Supramolecular protein cages constructed from a crystalline protein matrix (2018)
  • Author(s): Negishi Hashiru、Abe Satoshi、Yamashita Keitaro、Hirata Kunio、Niwase Kento、Boudes Marion、Coulibaly Fasseli、Mori Hajime、Ueno Takafumi
  • Journal Title: Chemical Communications Volume: 54 Issue: 16 Pages: 1988-1991
  • DOI: 10.1039/c7cc08689j
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Structure of in cell protein crystals containing organometallic complexes (2018)
  • Author(s): Abe Satoshi、Atsumi Kohei、Yamashita Keitaro、Hirata Kunio、Mori Hajime、Ueno Takafumi
  • Journal Title: Physical Chemistry Chemical Physics Volume: 20 Issue: 5 Pages: 2986-2989
  • DOI: 10.1039/c7cp06651a
  • Peer Reviewed
• [Journal Article] Photocatalytic hydrogen evolution systems constructed in cross-linked porous protein crystals (2018)
  • Author(s): Tabe Hiroyasu、Takahashi Hikaru、Shimoi Takuya、Abe Satoshi、Ueno Takafumi、Yamada Yusuke
  • Journal Title: Applied Catalysis B: Environmental Volume: 237 Pages: 1124-1129
  • DOI: 10.1016/j.apcatb.2018.01.046
  • URL: https://ocu-omu.repo.nii.ac.jp/records/2019618
  • Peer Reviewed
• [Journal Article] Artificial bio-nanomachines based on protein needles derived from bacteriophage T4 (2017)
  • Author(s): Inaba Hiroshi、Ueno Takafumi
  • Journal Title: Biophysical Reviews Volume: 10 Issue: 2 Pages: 641-658
  • DOI: 10.1007/s12551-017-0336-9
  • Peer Reviewed
• [Journal Article] Improved efficiency of nanoneedle insertion by modification with a cell-puncturing protein (2017)
  • Author(s): Ryu Seunghwan、Matsumoto Yuta、Matsumoto Takahiro、Ueno Takafumi、Silberberg Yaron R.、Nakamura Chikashi
  • Journal Title: Japanese Journal of Applied Physics Volume: 57 Issue: 3S2 Pages: 03EB02-03EB02
  • DOI: 10.7567/jjap.57.03eb02
  • NAID: 210000148743
  • Peer Reviewed
• [Journal Article] Development of Bio-Hybrid Materials by Design of Supramolecular Protein Assemblies (2017)
  • Author(s): 安部 聡、上野 隆史
  • Journal Title: Journal of Synthetic Organic Chemistry, Japan Volume: 75 Issue: 12 Pages: 1264-1273
  • DOI: 10.5059/yukigoseikyokaishi.75.1264
  • NAID: 130006248377
  • ISSN: 0037-9980, 1883-6526
  • Peer Reviewed
• [Journal Article] Observation of gold sub-nanocluster nucleation within a crystalline protein cage (2017)
  • Author(s): B. Maity, S. Abe, and T. Ueno
  • Journal Title: Nat. Commun. Volume: 8 Issue: 1 Pages: 14820-14825
  • DOI: 10.1038/ncomms14820
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Crystal Engineering of Self-Assembled Porous Protein Materials in Living Cells (2017)
  • Author(s): S. Abe, H. Tabe, H. Ijiri, K. Yamashita, K. Hirata, K. Atsumi, T. Shimoi, M. Akai, H. Mori, S. Kitagawa and T. Ueno
  • Journal Title: ACS Nano Volume: 11 Issue: 3 Pages: 2410-2419
  • DOI: 10.1021/acsnano.6b06099
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Design of Bioinorganic Materials At the Interface of Coordination and Biosupramolecular Chemistry (2017)
  • Author(s): B. Maity, and T. Ueno
  • Journal Title: Chem. Rec Volume: 17 Issue: 4 Pages: 383-398
  • DOI: 10.1002/tcr.201600122
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] Construction of Protein Needle and the Application for In Vivo MRI imaging (2018)
  • Author(s): Takafumi Ueno
  • Organizer: KAIST-Tokyo Tech Bio Joint Workshop
  • Int'l Joint Research / Invited
• [Presentation] Design of Protein Assembly for Supramolecular materials (2018)
  • Author(s): Takafumi Ueno
  • Organizer: Post-Mini workshop of Tokyo Tech Life Science and Technology International Symposium, “New Frontiers of Supramolecular Chemistry in Non-equilibrium Systems
  • Int'l Joint Research / Invited
• [Presentation] IN VIVO FUNCTIONS OF ORGANOMETALLOPROTEINS (2017)
  • Author(s): T Ueno
  • Organizer: 5th Symposium on Advanced Biological Inorganic Chemistry
  • Place of Presentation: Kolkata
  • Year and Date: 2017-01-08
  • Int'l Joint Research / Invited
• [Presentation] DESIGN OF PROTEIN ASSEMBLY FOR BIOINORGNIC MATERIALS (2017)
  • Author(s): Takafumi Ueno
  • Organizer: 11th Japan-China Joint Symposium on Metal Cluster Compounds
  • Int'l Joint Research / Invited
• [Presentation] Design of Bioinorganic Materials Constructed by Coordination Chemistry (2017)
  • Author(s): Takafumi Ueno
  • Organizer: 6th Asian Conference on Coordination Chemistry
  • Int'l Joint Research / Invited
• [Presentation] Functional Design of Protein Cage for Sustainable Bionanomaterial (2017)
  • Author(s): Takafumi Ueno
  • Organizer: ASPIRE FORUM 2017
  • Int'l Joint Research / Invited
• [Presentation] Design of Protein Assemblies as Molecular Templates (2017)
  • Author(s): Takafumi Ueno
  • Organizer: 10th China-Japan Joint Symposium on Functional Supramolecular Architectures
  • Int'l Joint Research / Invited
• [Presentation] Design of Protein Assemblies as Supramolecular Platforms (2017)
  • Author(s): Takafumi Ueno
  • Organizer: Frontier Bioorganization Forum 2017, “Dynamical ordering and integrated functions of biomolecular systems
  • Int'l Joint Research / Invited
• [Presentation] CO Releasing Bioinorganic Nanomaterials for CO Gas Biology (2017)
  • Author(s): Takafumi Ueno
  • Organizer: The International Advanced Drug Delivery Symposium 2017
  • Int'l Joint Research / Invited
• [Presentation] Design of Protein Asemblies in Development of Biohybrid Materials (2017)
  • Author(s): T Ueno
  • Organizer: Research Innovation Symposium in Life Science and Technology
  • Place of Presentation: Yokohama
  • Int'l Joint Research / Invited
• [Presentation] In Vivo and In Vitro Bioinorganic Functions Designed within Ferritin Cage” (2016)
  • Author(s): T Ueno
  • Organizer: 8th Asian Biological Inorganic Chemistry Conference
  • Place of Presentation: Auckland
  • Year and Date: 2016-12-06
  • Int'l Joint Research / Invited
• [Presentation] Multi-bioinorganic Functions Designed within Protein Cage (2016)
  • Author(s): T Ueno
  • Organizer: Japan-Korea-Taiwan Bioinorganic Chemistry Symposium 2016
  • Place of Presentation: Okazaki
  • Int'l Joint Research / Invited
• [Presentation] Biosupramolecular materials designed by protein assemblies (2016)
  • Author(s): T Ueno
  • Organizer: 2016 International Symposium on Polymer and Related Materials
  • Place of Presentation: Harbin
  • Int'l Joint Research / Invited
• [Book] フロンティア生物無機化学 (2016)
  • Author(s): 安部 聡、上野隆史
  • Total Pages: 350
  • Publisher: 三共出版
• [Remarks] The Ueno Lab
  • URL: http://www.ueno.bio.titech.ac.jp/
• [Remarks] タンパク質カゴの中で踊る金原子を観る
  • URL: http://educ.titech.ac.jp/bio/news/2017_03/053641.html
• [Remarks] タンパク質用いて細胞内分子フィルターを開発
  • URL: http://educ.titech.ac.jp/bio/news/2017_02/053432.html
• [Remarks] Ueno Laboratory
  • URL: http://www.ueno.bio.titech.ac.jp/
• [Patent(Industrial Property Rights)] 複合ポリペプチド単量体、細胞浸透機能を有する当該単量体の会合体、及び、当該会合体を有効成分とする免疫賦活剤 (2016)
  • Inventor(s): 上野隆史、吉川健吾、真野恵、片山和彦、三木元博、戸高 玲子
  • Industrial Property Rights Holder: 上野隆史、吉川健吾、真野恵、片山和彦、三木元博、戸高 玲子
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2016-207417
  • Filing Date: 2016-10-23
• [Patent(Industrial Property Rights)] 改変ポリヘドリンタンパク質及びその使用 (2016)
  • Inventor(s): 上野隆史、安部聡、笠松誠
  • Industrial Property Rights Holder: 上野隆史、安部聡、笠松誠
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2016-171201
  • Filing Date: 2016-09-01
• [Patent(Industrial Property Rights)] ヘモシアニン会合体を用いた包摂体の製造方法 (2016)
  • Inventor(s): 田中 良和、松野 明日香、北村 朗、上野 隆史、安部 聡
  • Industrial Property Rights Holder: 田中 良和、松野 明日香、北村 朗、上野 隆史、安部 聡
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2016-110309
  • Filing Date: 2016-06-01