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Calcium-dependent changes in intracellular phosphatidylserine distribution

Research Project

Project/Area Number 19K07265
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Review Section Basic Section 48010:Anatomy-related
Research InstitutionJuntendo University

Principal Investigator

Tsuji Takuma  順天堂大学, 大学院医学研究科, 特任助教 (40725628)

Project Period (FY) 2019-04-01 – 2023-03-31
Project Status Completed (Fiscal Year 2022)
Budget Amount *help
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2021: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2020: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2019: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Keywordsホスファチジルセリン / スクランブラーゼ / 電子顕微鏡 / 凍結割断レプリカ標識法 / 小胞体 / 凍結割断レプリカ法 / 膜脂質の非対称性分布 / 膜脂質の偏在性分布 / ホスファチジルイノシトール-(4,5)-二リン酸 / 膜脂質
Outline of Research at the Start

私達は新規に確立したホスファチジルセリン(PS)可視化法により、PSが小胞体膜の細胞質側に豊富に存在すること、細胞質Ca2+の上昇により小胞体膜内腔側にPSが出現し、同時に核膜PSが著しく増加すること等、新しい知見を得ました。本計画ではこれらをさらに発展させ、Ca2+依存的に細胞内PS分布が変化する分子機構、そしてその生物学的役割について研究します。さらに細胞内PS分布と疾病との関係を明らかにします。

Outline of Final Research Achievements

Phosphatidylserine (PS) is a major anionic phospholipid. Asymmetric distribution of PS in the plasma membrane is well established, and Ca2+-activated TMEM16F-dependent scrambling of PtdSer has been shown. In the present study, we developed a freeze-fracture electron microscopy method that can define the PS distribution in the individual leaflets of cellular membranes. Using this method, we found that the ER of mouse embryonic fibroblasts harbored abundant PS in the cytoplasmic leaflet and much less in the luminal leaflet. Significant amount of PS appeared in the luminal side of the ER upon treatment with the Ca2+ ionophore A23187 for 10 min. Interestingly, the A23187 treatment also increased PS in the nuclear membrane. This Ca2+-induced PS redistribution was not observed in TMEM16K KO cells, but was recovered in cells re-expressing Flag-TMEM16K. The results indicate that TMEM16K mediates Ca2+-dependent phospholipid scrambling in the ER.

Academic Significance and Societal Importance of the Research Achievements

細胞膜スクランブラーゼTMEM16F/ANO6は細胞内カルシウム濃度の上昇により活性化し、細胞質側のPSを細胞外側に輸送する。細胞内にもTMEM16ファミリー分子が存在し、全身性に発現するTMEM16K/ANO10は脊髄小脳変性症の原因遺伝子である。本研究で得られた成果は基礎生物学分野のみならず、医学的にも重要性が高いと言える。

Report

(5 results)
  • 2022 Annual Research Report   Final Research Report ( PDF )
  • 2021 Research-status Report
  • 2020 Research-status Report
  • 2019 Research-status Report
  • Research Products

    (24 results)

All 2022 2021 2020 2019 Other

All Int'l Joint Research (1 results) Journal Article (9 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 9 results,  Open Access: 5 results) Presentation (12 results) (of which Int'l Joint Research: 2 results,  Invited: 10 results) Book (2 results)

  • [Int'l Joint Research] Center for Molecular Biology/Heidelberg University(ドイツ)

    • Related Report
      2019 Research-status Report
  • [Journal Article] Distribution of Membrane Lipids Revealed by TEM2022

    • Author(s)
      Tsuji Takuma
    • Journal Title

      MEMBRANE

      Volume: 47 Issue: 5 Pages: 281-285

    • DOI

      10.5360/membrane.47.281

    • ISSN
      0385-1036, 1884-6440
    • Related Report
      2022 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Ultrastructural localization of de novo synthesized phosphatidylcholine in yeast cells by freeze-fracture electron microscopy2021

    • Author(s)
      Tsuji Takuma、Fujimoto Toyoshi
    • Journal Title

      STAR Protocols

      Volume: 2 Issue: 4 Pages: 100990-100990

    • DOI

      10.1016/j.xpro.2021.100990

    • Related Report
      2021 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Characterization of micron-scale protein-depleted plasma membrane domains in phosphatidylserine-deficient yeast cells2021

    • Author(s)
      Mioka Tetsuo、Guo Tian、Wang Shiyao、Tsuji Takuma、Kishimoto Takuma、Fujimoto Toyoshi、Tanaka Kazuma
    • Journal Title

      Journal of Cell Science

      Volume: 135 Issue: 5

    • DOI

      10.1242/jcs.256529

    • Related Report
      2021 Research-status Report
    • Peer Reviewed
  • [Journal Article] Transmembrane phospholipid translocation mediated by Atg9 is involved in autophagosome formation2021

    • Author(s)
      Orii Minami、Tsuji Takuma、Ogasawara Yuta、Fujimoto Toyoshi
    • Journal Title

      Journal of Cell Biology

      Volume: 220 Issue: 3

    • DOI

      10.1083/jcb.202009194

    • Related Report
      2020 Research-status Report
    • Peer Reviewed
  • [Journal Article] Atg9 is a lipid scramblase that mediates autophagosomal membrane expansion2020

    • Author(s)
      Matoba Kazuaki、Kotani Tetsuya、Tsutsumi Akihisa、Tsuji Takuma、Mori Takaharu、Noshiro Daisuke、Sugita Yuji、Nomura Norimichi、Iwata So、Ohsumi Yoshinori、Fujimoto Toyoshi、Nakatogawa Hitoshi、Kikkawa Masahide、Noda Nobuo N.
    • Journal Title

      Nature Structural & Molecular Biology

      Volume: 27 Issue: 12 Pages: 1185-1193

    • DOI

      10.1038/s41594-020-00518-w

    • Related Report
      2020 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Multifarious roles of lipid droplets in autophagy ? Target, product, and what else?2020

    • Author(s)
      Ogasawara Yuta、Tsuji Takuma、Fujimoto Toyoshi
    • Journal Title

      Seminars in Cell & Developmental Biology

      Volume: - Pages: 47-54

    • DOI

      10.1016/j.semcdb.2020.02.013

    • Related Report
      2020 Research-status Report
    • Peer Reviewed
  • [Journal Article] Predominant localization of phosphatidylserine at the cytoplasmic leaflet of the ER, and its TMEM16K-dependent redistribution2019

    • Author(s)
      Tsuji Takuma、Cheng Jinglei、Tatematsu Tsuyako、Ebata Aoi、Kamikawa Hiroki、Fujita Akikazu、Gyobu Sayuri、Segawa Katsumori、Arai Hiroyuki、Taguchi Tomohiko、Nagata Shigekazu、Fujimoto Toyoshi
    • Journal Title

      Proceedings of the National Academy of Sciences

      Volume: 116 Issue: 27 Pages: 13368

    • DOI

      10.1073/pnas.1822025116

    • URL

      https://pure.teikyo.jp/en/publications/e62284b2-5567-40aa-b36c-45c5ca281a73

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] ESCRT machinery mediates selective microautophagy of endoplasmic reticulum in yeast2019

    • Author(s)
      Schafer Jasmin A、Schessner Julia P、Bircham Peter W、Tsuji Takuma、Funaya Charlotta、Pajonk Oliver、Schaeff Katharina、Ruffini Giulia、Papagiannidis Dimitrios、Knop Michael、Fujimoto Toyoshi、Schuck Sebastian
    • Journal Title

      The EMBO Journal

      Volume: 39 Issue: 2

    • DOI

      10.15252/embj.2019102586

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] A method to selectively internalise submicrometer boron carbide particles into cancer cells using surface transferrin conjugation for developing a new boron neutron capture therapy agent2019

    • Author(s)
      Tsuji Takuma、Yoshitomi Hiroshi、Ishikawa Yoshie、Koshizaki Naoto、Suzuki Motoshi、Usukura Jiro
    • Journal Title

      Journal of Experimental Nanoscience

      Volume: 15 Issue: 1 Pages: 1-11

    • DOI

      10.1080/17458080.2019.1692178

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] オルガネラ形態変化とミクロリポファジー2022

    • Author(s)
      辻琢磨
    • Organizer
      第45回日本分子生物学会年会
    • Related Report
      2022 Annual Research Report
    • Invited
  • [Presentation] Molecular mechanism of micro-lipophagy in yeast2022

    • Author(s)
      Tsuji T
    • Organizer
      72th Annual Meeting Korean Association of Anatomists
    • Related Report
      2022 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 電子顕微鏡による膜脂質分布の解析2022

    • Author(s)
      辻琢磨, 藤本豊士
    • Organizer
      日本膜学会第44年会
    • Related Report
      2022 Annual Research Report
    • Invited
  • [Presentation] Molecular mechanism of micro-lipophagy in budding yeas2022

    • Author(s)
      Tsuji T, Fujimoto T
    • Organizer
      日本顕微鏡学会第78回学術講演会
    • Related Report
      2022 Annual Research Report
    • Invited
  • [Presentation] 出芽酵母におけるミクロリポファジー機構の解析2022

    • Author(s)
      辻琢磨, 藤本豊士
    • Organizer
      第127回日本解剖学会総会・全国学術集会
    • Related Report
      2021 Research-status Report
    • Invited
  • [Presentation] Distribution of vacuolar proteins in autophagy deficient yeast2021

    • Author(s)
      Tsuji T, Shibata R, Fujimoto T
    • Organizer
      第44回日本分子生物学会年会
    • Related Report
      2021 Research-status Report
    • Invited
  • [Presentation] 電子顕微鏡による膜脂質の解析2021

    • Author(s)
      辻琢磨、藤本豊士
    • Organizer
      第126回日本解剖学会総会・全国学術集会 第98回日本生理学会大会 合同大会
    • Related Report
      2020 Research-status Report
    • Invited
  • [Presentation] 膜脂質の対称性・非対称性分布2021

    • Author(s)
      辻琢磨、藤本豊士
    • Organizer
      日本顕微鏡学会 第45回関東支部講演会
    • Related Report
      2020 Research-status Report
    • Invited
  • [Presentation] 電子顕微鏡による膜脂質分子の可視化2020

    • Author(s)
      辻琢磨
    • Organizer
      北海道大学 先端生命科学研究院 細胞装置学セミナー
    • Related Report
      2020 Research-status Report
    • Invited
  • [Presentation] 電子顕微鏡による膜脂質局在解析2020

    • Author(s)
      辻琢磨、藤本豊士
    • Organizer
      第93回日本生化学会大会
    • Related Report
      2020 Research-status Report
    • Invited
  • [Presentation] 形質膜のホスファチジルセリンとカベオラ2020

    • Author(s)
      辻琢磨、藤本豊士
    • Organizer
      第125回日本解剖学会総会・全国学術集会
    • Related Report
      2019 Research-status Report
  • [Presentation] Ca2+-dependent phosphatidylserine redistribution in the ER2019

    • Author(s)
      Tsuji T, Cheng J, Tatematsu T, Ebata A, Kamikawa H, Taguchi T, Fujimoto T
    • Organizer
      60th International Conference on the Bioscience of Lipids
    • Related Report
      2019 Research-status Report
    • Int'l Joint Research
  • [Book] The Lipid 小胞体のリン脂質分布制御機構と関連する疾患2021

    • Author(s)
      辻琢磨, 藤本豊士
    • Total Pages
      7
    • Publisher
      メディカルレビュー社
    • Related Report
      2021 Research-status Report
  • [Book] 脂質解析ハンドブック 脂質を解析する技術 電子顕微鏡を用いた観察2019

    • Author(s)
      辻琢磨、藤本豊士
    • Total Pages
      8
    • Publisher
      羊土社
    • ISBN
      9784758122412
    • Related Report
      2019 Research-status Report

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Published: 2019-04-18   Modified: 2024-01-30  

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