Evaluating the affinity of membrane proteins and lipid raft using a patterned model membrane platform
| Project/Area Number |
17H03666
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Biophysics
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| Research Institution | Kobe University |
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Principal Investigator |
Morigaki Kenichi 神戸大学, バイオシグナル総合研究センター, 准教授 (10358179)
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| Co-Investigator(Kenkyū-buntansha) |
林 文夫 神戸大学, 理学研究科, 名誉教授 (80093524)
鈴木 健一 岐阜大学, 研究推進・社会連携機構, 教授 (50423059)
笠井 倫志 京都大学, ウイルス・再生医科学研究所, 助教 (20447949)
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| Project Period (FY) |
2017-04-01 – 2020-03-31
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| Project Status |
Completed (Fiscal Year 2019)
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| Budget Amount *help |
¥17,550,000 (Direct Cost: ¥13,500,000、Indirect Cost: ¥4,050,000)
Fiscal Year 2019: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2018: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2017: ¥8,580,000 (Direct Cost: ¥6,600,000、Indirect Cost: ¥1,980,000)
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| Keywords | 生体膜 / ラフト / 受容体 / 人工膜 / 蛍光顕微鏡 / 膜小胞 / 脂質ラフト / 膜タンパク質 / Gタンパク質共役型受容体 / GPIアンカー型タンパク質 |
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| Outline of Final Research Achievements |
Model biological membranes are useful tools to study the functions of membrane proteins. We developed a strategy to directly reconstitute mammalian membrane proteins from the cell membrane blebs into a model biological membrane to bypass the technically challenging solubilization and purification processes. We expressed GPCR and GPI-AP in CHO cells and produced blebs by chemical induction. By rupturing blebs on the substrate surface, we could form a planar bilayer and observe single molecules of GPCR and GPI-AP. Furthermore, many reconstituted molecules were observed in a nanometric cleft between the substrate and a PDMS elastomer sheet. This methodology should enable to evaluate the physicochemical properties and functions for a wide range of mammalian membrane proteins.
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| Academic Significance and Societal Importance of the Research Achievements |
膜タンパク質は、細胞内においてシグナル伝達など重要な機能を担い、疾患などにも深く関与しているため、医薬品の重要な標的になっている。膜タンパク質の機能は脂質膜の組成や分布にも影響を受けていると考えらてるが、生体膜内における機能制御機構を生きた細胞を用いて詳細に検証することは非常に困難である。本研究により、哺乳類膜タンパク質を細胞から人工膜に導入する技術が開発され、その機能解析に道が開けた。この成果は、膜タンパク質の機能を分子レベルで定量的に評価することを可能にし、疾患メカニズムの解明、医薬品開発にも貢献できるものと考えられる。
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Report
(4 results)
Research Products
(95 results)
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[Journal Article] Defining raft domains in the plasma membrane2019
Author(s)
Akihiro Kusumi、Takahiro K. Fujiwara、Taka A. Tsunoyama、Rinshi S. Kasai、An‐An Liu、Koichiro M. Hirosawa、Masanao Kinoshita、Nobuaki Matsumori、Naoko Komura、Hiromune Ando、Kenichi G. N. Suzuki
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Journal Title
Traffic
Volume: 21
Issue: 1
Pages: 106-137
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research
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[Journal Article] The role of the Prod1 membrane anchor in newt limb regeneration2017
Author(s)
Nomura, K., Tanimoto, Y., Hayashi, F., Harada, E., Shan, X.-Y., Shionyu, M., Hijikata, A., Shirai, T., Morigaki, K., Shimamoto, K.
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Journal Title
Angew. Chem. Int. Ed.
Volume: 56
Issue: 1
Pages: 270-274
DOI
Related Report
Peer Reviewed / Open Access
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[Journal Article] Raft-based sphingomyelin interactions revealed by new fluorescent sphingomyelin analogs2017
Author(s)
Masanao Kinoshita, Kenichi GN Suzuki, Nobuaki Matsumori, Misa Takada, Hikaru Ano, Kenichi Morigaki, Mitsuhiro Abe, Asami Makino, Toshihide Kobayashi, Koichiro M Hirosawa, Takahiro K Fujiwara, Akihiro Kusumi, Michio Murata
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Journal Title
J Cell Biol
Volume: 未定
Issue: 4
Pages: 1183-1204
DOI
NAID
Related Report
Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
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[Presentation] Hybrid model membrane combining micropatterned lipid bilayer and hydrophilic polymer brush2017
Author(s)
Morigaki, K., Nishimura, T., Tamura, F., Kobayashi, S., Tanimoto, Y., Sudo, Y., Hayashi, F., Iwasaki, K.
Organizer
253rd American Chemical Society National Meeting
Related Report
Int'l Joint Research / Invited
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