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Artificial cell systems for modeling of cell-information cross-talk and cell dynamic analysis

Research Project

Project/Area Number 16K07293
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Functional biochemistry
Research InstitutionMie University

Principal Investigator

Tsumoto Kanta  三重大学, 工学研究科, 准教授 (80362359)

Co-Investigator(Kenkyū-buntansha) 瀧口 金吾  名古屋大学, 理学研究科, 講師 (20262842)
Project Period (FY) 2016-04-01 – 2020-03-31
Project Status Completed (Fiscal Year 2019)
Budget Amount *help
¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000)
Fiscal Year 2018: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000)
Fiscal Year 2017: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2016: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Keywords人工細胞 / 細胞情報 / プロテオリポソーム / 膜タンパク質 / 細胞骨格 / 脂質二分子膜 / GUV / マイクロコンパートメント / 人工膜 / リポソーム
Outline of Final Research Achievements

In this project, we tried to study and develop how to incorporate proteins related to cell signal transduction and cell motility into artificial lipid bilayer systems mimicking actual membranes of living cells. In addition, through realization of such model systems, we would like to believe we can invent a novel analytical protocol for artificial cellular dynamics. In this study, we demonstrated the following results: reconstitution of recombinant membrane proteins like GPCRs into giant unilamellar vesicles (GUVs) through membrane fusion with baculovirus budded virions (BVs), high-yield preparation of GUVs with high density, and apparently selective accumulation of the biomacromolecules cytoskeletal proteins like filamentous actin into aqueous microcompartments.

Academic Significance and Societal Importance of the Research Achievements

複雑な細胞機能を、現代のオープンバイオリソースを活用しながらモデル化された実験系の上に実現することは、細胞生命現象の別角度からの理解に役立つとともに、そこで得た知見が工学的なイノベーションの種となるとも、期待される。細胞内では多種多様なプロセスが併存して機能しているが、研究に人工細胞モデルを用いる意義は、シンプルな経路として取り出すことのほか、プロセスの並行を可能とする本質に、眼前で実現することで実験科学的に迫ることである。今回、人工細胞膜へ組換え膜受容体を構成できること、多量高密度の人工細胞ベシクルを広い条件で作れること、細胞骨格タンパク質を高濃度で区画に封じ込めること、をそれぞれ実証した。

Report

(5 results)
  • 2019 Annual Research Report   Final Research Report ( PDF )
  • 2018 Research-status Report
  • 2017 Research-status Report
  • 2016 Research-status Report
  • Research Products

    (44 results)

All 2019 2018 2017 2016 Other

All Journal Article (7 results) (of which Peer Reviewed: 7 results,  Open Access: 4 results) Presentation (33 results) (of which Int'l Joint Research: 13 results,  Invited: 5 results) Remarks (4 results)

  • [Journal Article] Membrane fusion and infection abilities of baculovirus virions are preserved during freezing and thawing in the presence of trehalose2019

    • Author(s)
      Nakanishi Kohei、Tomita Masahiro、Tsumoto Kanta
    • Journal Title

      Bioscience, Biotechnology, and Biochemistry

      Volume: 84 Issue: 4 Pages: 686-694

    • DOI

      10.1080/09168451.2019.1704396

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Aqueous/Aqueous Micro Phase Separation: Construction of an Artificial Model of Cellular Assembly2019

    • Author(s)
      Sakuta Hiroki、Fujimoto Tadashi、Yamana Yusuke、Hoda Yusuke、Tsumoto Kanta、Yoshikawa Kenichi
    • Journal Title

      Frontiers in Chemistry

      Volume: 7 Pages: 44-44

    • DOI

      10.3389/fchem.2019.00044

    • Related Report
      2018 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Specific Spatial Localization of Actin and DNA in a?Water/Water Microdroplet: Self-Emergence of a Cell-Like Structure2018

    • Author(s)
      Nakatani Naoki、Sakuta Hiroki、Hayashi Masahito、Tanaka Shunsuke、Takiguchi Kingo、Tsumoto Kanta、Yoshikawa Kenichi
    • Journal Title

      ChemBioChem

      Volume: 19 Issue: 13 Pages: 1370-1374

    • DOI

      10.1002/cbic.201800066

    • Related Report
      2018 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Repetitive stretching of giant liposomes utilizing the nematic alignment of confined actin2018

    • Author(s)
      Tanaka Shunsuke、Takiguchi Kingo、Hayashi Masahito
    • Journal Title

      Communications Physics

      Volume: 1 Issue: 1 Pages: 18-18

    • DOI

      10.1038/s42005-018-0019-2

    • Related Report
      2018 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] A reverse-phase method revisited: Rapid high-yield preparation of giant unilamellar vesicles (GUVs) using emulsification followed by centrifugation2018

    • Author(s)
      Tsumoto Kanta、Hayashi Yuki、Tabata Jin、Tomita Masahiro
    • Journal Title

      Colloids and Surfaces A: Physicochemical and Engineering Aspects

      Volume: 546 Pages: 74-82

    • DOI

      10.1016/j.colsurfa.2018.02.060

    • Related Report
      2017 Research-status Report
    • Peer Reviewed
  • [Journal Article] Membrane fusion between baculovirus budded virus-enveloped particles and giant liposomes generated using a droplet-transfer method for the incorporation of recombinant membrane proteins2017

    • Author(s)
      Nishigami Misako、Mori Takaaki、Tomita Masahiro、Takiguchi Kingo、Tsumoto Kanta
    • Journal Title

      Colloids and Surfaces B: Biointerfaces

      Volume: 155 Pages: 248-256

    • DOI

      10.1016/j.colsurfb.2017.04.027

    • Related Report
      2017 Research-status Report
    • Peer Reviewed
  • [Journal Article] The Aqueous Two Phase System (ATPS) Deserves Plausible Real-World Modeling for the Structure and Function of Living Cells2017

    • Author(s)
      Tsumoto Kanta、Yoshikawa Kenichi
    • Journal Title

      MRS Advances

      Volume: 2 Issue: 45 Pages: 2407-2413

    • DOI

      10.1557/adv.2017.358

    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] 組換えバキュロウイルス膜融合によるプロテオGUV形成の効率化2019

    • Author(s)
      伊藤諒,中西航平,冨田昌弘,湊元幹太
    • Organizer
      第83回日本生化学会中部支部例会・シンポジウム
    • Related Report
      2019 Annual Research Report
  • [Presentation] 組換えバキュロウイルス膜融合によるプロテオGUV形成の効率化と膜機能解析2019

    • Author(s)
      伊藤諒, 中西航平, 冨田昌弘, 湊元幹太
    • Organizer
      第92回日本生化学会大会
    • Related Report
      2019 Annual Research Report
  • [Presentation] Baculovirus particles for displaying/delivering recombinant membrane proteins onto artificial lipid bilayers2019

    • Author(s)
      K. Tsumoto, S. Nishio, H. Seto, K. Nakanishi, M. Tomita
    • Organizer
      13th International Symposium on Nanomedicine (ISNM2019)
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Giant vesicles rapidly prepared and purified using a reverse-phase/centrifugation method2018

    • Author(s)
      Kanta Tsumoto, Kohei Nakano, Yuki Hayashi, Masahiro Tomita
    • Organizer
      第56回日本生物物理学会年会
    • Related Report
      2018 Research-status Report
  • [Presentation] 界面通過法により作製したGUVによるカルシウムイオン膜透過性評価2018

    • Author(s)
      宇野勇気,西上美佐子,冨田昌弘,湊元幹太
    • Organizer
      第91回日本生化学会大会
    • Related Report
      2018 Research-status Report
  • [Presentation] Cell-Sized Lipid Vesicles as Microcompartments for Mimicking Cells: Rapid Prepartion and Installation of Recombinant Proteins2018

    • Author(s)
      Kanta Tsumoto, Kohei Nakanishi, Yuki Hayashi, Masahiro Tomita
    • Organizer
      The 6th Japan-China Symposimu on Nanomedicine
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] 逆相遠心法による細胞サイズベシクル調製と膜ミクロドメイン形成2018

    • Author(s)
      仲野耕平,林佑紀,冨田昌弘,湊元幹太
    • Organizer
      第82回日本生化学会中部支部例会・シンポジウム
    • Related Report
      2018 Research-status Report
  • [Presentation] Monitoring of Calcium Ion Permeation Using Single GUVs Prepared with the Droplet Transfer Method2018

    • Author(s)
      Y. Uno, M. Nishigami, M. Tomita, K. Tsumoto
    • Organizer
      12th International Symposium on Nanomedicine
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research
  • [Presentation] バキュロウイルス懸濁液におけるトレハロースの凍結保護効果:膜融合・感染能・形態の比較から2018

    • Author(s)
      中西 航平, 冨田 昌弘, 湊元 幹太
    • Organizer
      第91回日本生化学会大会
    • Related Report
      2018 Research-status Report
  • [Presentation] Prepartion of ProteoSS-BLMs Using the Membrane Fusion between Recombinant Baculovirus and SS-BLMs2018

    • Author(s)
      K. Nakanishi, M. Tomita and K. Tsumoto
    • Organizer
      12th International Symposium on Nanomedicine
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research
  • [Presentation] 細胞骨格封入リポソームの屈伸運動2018

    • Author(s)
      林真人、田中駿介、西山雅祥、瀧口金吾
    • Organizer
      第1回分子ロボティクス年次大会(併催分子ロボット倫理シンポジウム)
    • Related Report
      2017 Research-status Report
  • [Presentation] ATPS Deserves Plausible Real-World Modeling for the Structure and Function of Living Cells2017

    • Author(s)
      Kanta Tsumoto
    • Organizer
      2017 MRS (Materials Research Society) Spring Meeting & Exhibit, Symposium SM5 : Aqueous Cytomimetic Materials
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] 組換えバキュロウイルスを用いた膜タンパク質包埋型球状支持二重膜(SS-BLM)の調製2017

    • Author(s)
      中西航平,西上美佐子,冨田昌弘,湊元幹太
    • Organizer
      第81回日本生化学会中部支部例会・シンポジウム
    • Related Report
      2017 Research-status Report
  • [Presentation] 逆相遠心法による巨大リン脂質ベシクルの効率作製:安定性、物質保持、膜ドメインと生理条件での生成について2017

    • Author(s)
      林佑紀,冨田昌弘,湊元幹太
    • Organizer
      第81回日本生化学会中部支部例会・シンポジウム
    • Related Report
      2017 Research-status Report
  • [Presentation] Analysis of membrane fusion between membrane protein-expressing baculovirus virions and GUVs prepared with a droplet transfer method2017

    • Author(s)
      西上 美佐子, 冨田 昌弘, 瀧口 金吾, 湊元 幹太
    • Organizer
      第55回日本生物物理学会年会
    • Related Report
      2017 Research-status Report
  • [Presentation] Preparation of ProteoSS-BLM (Spherically Supported Bilayer Membrane) Using Recombinant Baculovirus2017

    • Author(s)
      Kohei Nakanishi, Masahiro Tomita, Kanta Tsumoto
    • Organizer
      IS2EMU 2017 The 7th International Symposium for Sustainability by Engineering at MIU Research Area D & F Life Science &Advanced Materials
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research
  • [Presentation] Large-Scale Preparation of Biocompatible GUVs Using a Reverse Phase/Centrifugation Method2017

    • Author(s)
      Yuki Hayashi, Kohei Nakano, Masahiro Tomita, Kanta Tsumoto
    • Organizer
      IS2EMU 2017 The 7th International Symposium for Sustainability by Engineering at MIU Research Area D & F Life Science &Advanced Materials
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research
  • [Presentation] Effects of Coexisting/Entrapped Proteins on GUVs Prepared with the Reverse Phase/Centrifugation Method2017

    • Author(s)
      Kohei Nakano, Yuki Hayashi, Masahiro Tomita, Kanta Tsumoto
    • Organizer
      IS2EMU 2017 The 7th International Symposium for Sustainability by Engineering at MIU Research Area D & F Life Science &Advanced Materials
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research
  • [Presentation] 逆相遠心法による巨大リン脂質ベシクルの形成と高効率精製2017

    • Author(s)
      林 佑紀、仲野 耕平、冨田 昌弘、湊元 幹太
    • Organizer
      ConBio2017 (2017年度 生命科学系学会合同年次大会) 第90回日本生化学会大会
    • Related Report
      2017 Research-status Report
  • [Presentation] 界面通過法を用いた細胞サイズリポソーム-バキュロウイルス粒子の膜融合解析と組換え膜タンパク質の組込み2017

    • Author(s)
      西上 美佐子、冨田 昌弘、瀧口 金吾、湊元 幹太
    • Organizer
      ConBio2017 (2017年度 生命科学系学会合同年次大会) 第90回日本生化学会大会
    • Related Report
      2017 Research-status Report
  • [Presentation] 組換えバキュロウイルスを利用した球状支持二重膜(SS-BLM)への膜タンパク質再構成法の検討2017

    • Author(s)
      中西 航平、冨田 昌弘、湊元 幹太
    • Organizer
      ConBio2017 (2017年度 生命科学系学会合同年次大会) 第90回日本生化学会大会
    • Related Report
      2017 Research-status Report
  • [Presentation] Preparation of proteoSS-BLM (spherically supported bilayer membrane) using recombinant baculovirus virions2017

    • Author(s)
      Kohei Naknishi, Masahiro Tomita, Kanta Tsumoto
    • Organizer
      11th International Symposium on Nanomedicine (ISNM2017)
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research
  • [Presentation] Simple Preparation of Giant Liposomes for Mimicking Cell Systems2017

    • Author(s)
      K. Tsumoto, Y. Hayashi, K. Nakano, M. Tomita
    • Organizer
      11th International Symposium on Nanomedicine (ISNM2017)
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] 細胞サイズを持つ巨大リポソームの細胞骨格による形態制御 --- 変形・運動を繰り返し行う人工細胞の構築に向けて ---2017

    • Author(s)
      瀧口金吾 田中駿介 林真人
    • Organizer
      新学術領域「分子ロボティクス」最終公開シンポジウム 「分子ロボティクスの拓く未来」
    • Related Report
      2017 Research-status Report
  • [Presentation] Light-induced and reversible morphological control of F-actin-encapsulating giant liposomes2017

    • Author(s)
      Hayashi M., Tanaka S., Takiguchi K
    • Organizer
      第55回日本生物物理学会年会
    • Related Report
      2017 Research-status Report
  • [Presentation] 細胞サイズを持つ巨大リポソームの細胞骨格による形態制御2017

    • Author(s)
      瀧口金吾 田中駿介 林真人
    • Organizer
      第7回ソフトマター研究会
    • Related Report
      2017 Research-status Report
  • [Presentation] Preparation and purification of GUVs with a reverse-phase method2016

    • Author(s)
      Yuki Hayashi, Masahiro Tomita, Kanta Tsumoto
    • Organizer
      10th Anniversary Internationla Symposium on Nanomedicine (ISNM2016)
    • Place of Presentation
      Auditorium of National Institute of Advanced Industrial Science and Technology (AIST) (Tsukuba-shi)
    • Year and Date
      2016-11-24
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research
  • [Presentation] Visualization of membrane fusion between multicomponent protein-expressing BVs and GUVs prepared using a droplet-transfer method2016

    • Author(s)
      Misako Nishigami, Masahiro Tomita, Kanta Tsumoto
    • Organizer
      10th Anniversary Internationla Symposium on Nanomedicine (ISNM2016)
    • Place of Presentation
      Auditorium of National Institute of Advanced Industrial Science and Technology (AIST) (Tsukuba-shi)
    • Year and Date
      2016-11-24
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research
  • [Presentation] インテグリンを再構成した巨大組換えプロテオリポソームの調製2016

    • Author(s)
      中西航平, 西上美佐子, 冨田昌弘, 湊元幹太
    • Organizer
      第89回日本生化学会大会
    • Place of Presentation
      仙台国際センター/東北大学川内北キャンパス(仙台市)
    • Year and Date
      2016-09-25
    • Related Report
      2016 Research-status Report
  • [Presentation] Recombinant Baculovirus Particles as Resources of Membrane Proteins for Artificial Cell Membrane Reconstitution2016

    • Author(s)
      Kanta Tsumoto, Kohei Nakanishi, Tomomi Hattori, Masahiro Tomita
    • Organizer
      The Fourth Japan-Chaina Symposium on Nanomedicine
    • Place of Presentation
      Kitakyushu International Conference Center(Kitakyushu-shi)
    • Year and Date
      2016-05-12
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] 逆相法による巨大リン脂質ベシクルの形成と内外相の比重差を利用した単離2016

    • Author(s)
      林佑紀,冨田昌弘,湊元幹太
    • Organizer
      第80回日本生化学会中部支部例会・シンポジウム
    • Place of Presentation
      三重大学講堂(津市)
    • Related Report
      2016 Research-status Report
  • [Presentation] 組換えバキュロウイルスを用いたインテグリンプロテオリポソーム調製手法の検討2016

    • Author(s)
      中西航平,西上美佐子,冨田昌弘,湊元幹太
    • Organizer
      第80回日本生化学会中部支部例会・シンポジウム
    • Place of Presentation
      三重大学講堂(津市)
    • Related Report
      2016 Research-status Report
  • [Presentation] バキュロウイルス膜融合法を用いた膜タンパク質再構成リポソームの作製と人工細胞モデルの構築2016

    • Author(s)
      中西航平、西上美佐子、冨田昌弘、湊元幹太
    • Organizer
      第6回地域イノベーション学会大会
    • Place of Presentation
      三重大学(津市)
    • Related Report
      2016 Research-status Report
  • [Remarks] Google Scholar Citations

    • URL

      https://scholar.google.com/citations?user=HesLhbwAAAAJ&hl=ja

    • Related Report
      2019 Annual Research Report 2018 Research-status Report
  • [Remarks] Google Scholar Citations

    • URL

      https://scholar.google.com/citations?hl=ja&user=HesLhbwAAAAJ

    • Related Report
      2017 Research-status Report 2016 Research-status Report
  • [Remarks] researchmap

    • URL

      http://researchmap.jp/read0067280/

    • Related Report
      2017 Research-status Report 2016 Research-status Report
  • [Remarks] 分子生物工学研究室

    • URL

      http://www.bio.chem.mie-u.ac.jp/

    • Related Report
      2017 Research-status Report 2016 Research-status Report

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Published: 2016-04-21   Modified: 2021-02-19  

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