Development of sodium-selective optogenetic tools for super-precise optical neural manipulation
| Project/Area Number |
20K21383
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| Research Category |
Grant-in-Aid for Challenging Research (Exploratory)
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| Allocation Type | Multi-year Fund |
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| Review Section |
Medium-sized Section 43:Biology at molecular to cellular levels, and related fields
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| Research Institution | The University of Tokyo |
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Principal Investigator |
Inoue Keiichi 東京大学, 物性研究所, 准教授 (90467001)
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| Project Period (FY) |
2020-07-30 – 2023-03-31
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| Project Status |
Completed (Fiscal Year 2022)
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| Budget Amount *help |
¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2022: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2021: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2020: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
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| Keywords | ロドプシン / レチナール / オプトジェネティクス / ナトリウムポンプ / ヘリオロドプシン |
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| Outline of Research at the Start |
本研究では近年、光で神経興奮を操作するオプトジェネティクスが大きな注目を集めている、光駆動型イオン輸送膜タンパク質の微生物ロドプシンを用いて、Na+選択的なオプトジェネティクスツールの開発を行う。
そのため、最近研究代表者らが発見した外向きNa+ポンプ型ロドプシン（NaR）とヘリオロドプシン（HeR）の二種類の微生物型ロドプシンをもとに、Na+のみを高効率で細胞内へ輸送するロドプシンの開発を行う。さらに得られた分子を神経細胞に導入して、Na+選択輸送による神経活動の光操作を実現する。
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| Outline of Final Research Achievements |
In this study, we aimed to create an inwardly oriented sodium pumping rhodopsin by reversing the orientation of natural outward sodium pumping rhodopsin in the membrane, thereby selectively transporting only sodium ions into cells for the development of optogenetic tools. This was accomplished by altering the distribution of charged residues on the extracellular and intracellular surfaces of the outward sodium pumping rhodopsin. To achieve this, we used the BEdep_AAProf method to design a molecule with a reversed charge distribution, which presented numerous mutation sites for causing orientation reversal. Experimental results using E. coli demonstrated that it is feasible to stably introduce eight to nine mutations. This significant finding provides a technological foundation for future development of optogenetic tools.
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| Academic Significance and Societal Importance of the Research Achievements |
本研究ではBEdep_AAProf法を用いることで、計算科学的に電荷部分布を変えた安定なタンパク質分子を設計可能であることが示された。これは今後、医療および産業面において重要な機能性分子の開発にも新たな方法論を与えるものである。また実験で得られた外向きナトリウムポンプ型ロドプシンの８～９重変異体は、今後N末に膜貫通ヘリックスを追加することで配向を反転させたロドプシンの構造をさらに安定化することで、ナトリウムイオン選択的なオプトジェネティクスツールの開発に貢献すると期待される。
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Report
(4 results)
Research Products
(119 results)
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[Journal Article] Phototrophy by Antenna-Containing Rhodopsin Pumps in Aquatic Environments2023
Author(s)
Chazan, Das, Fujiwara, Murakoshi, Rozenberg, Molina-Marquez, Sano, Tanaka, Gomez-Villegas, Larom, Pushkarev, Malakar, Hasegawa, Tsukamoto, Ishizuka, Konno, Nagata, Mizuno, Katayama, Abe-Yoshizumi, Ruhman, Inoue, Kandori, Leon, Shihoya, Yoshizawa, Sheves, Nureki, Beja
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Journal Title
Nature
Volume: 615
Issue: 7952
Pages: 535-540
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research
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[Journal Article] Rhodopsin-bestrophin fusion proteins from unicellular algae form gigantic pentameric ion channels2022
Author(s)
Rozenberg A,Kaczmarczyk I,Matzov D,Vierock J,Nagata T,Sugiura M,Katayama K,Kawasaki Y,Konno M,Nagasaka Y,Aoyama M,Das I,Pahima E,Church J,Adam S,Borin VA,Chazan A,Augustin S,Wietek J,Dine J,Peleg Y,Kawanabe A,Fujiwara Y,Yizhar O,Sheves M,Schapiro I,Furutani Y,Kandori H,Inoue K,Hegemann P,Beja O,Shalev-Benami M.
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Journal Title
Nature Structural & Molecular Biology
Volume: 29
Issue: 6
Pages: 592-603
DOI
Related Report
Peer Reviewed / Int'l Joint Research
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[Journal Article] Diverse Heliorhodopsins Detected via Functional Metagenomics in Freshwater Actinobacteria, Chloroflexi and Archaea2022
Author(s)
Ariel Chazan, Andrey Rozenberg, Kentaro Mannen, Takashi Nagata, Ran Tahan, Shir Yaish, Shirley Larom, Keiichi Inoue, Oded Beja, Alina Pushkarev
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Journal Title
Environmental Microbiology
Volume: 24
Issue: 1
Pages: 110-121
DOI
Related Report
Peer Reviewed / Int'l Joint Research
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[Journal Article] Schizorhodopsins: A novel family of rhodopsins from Asgard archaea that function as light-driven inward H+ pumps2020
Author(s)
Keiichi Inoue, Satoshi P. Tsunoda, Manish Singh, Sahoko Tomida, Shoko Hososhima, Masae Konno, Ryoko Nakamura, Hiroki Watanabe, Paul-Adrian Bulzu, Horia L. Banciu, Adrian Andrei, Takayuki Uchihashi, Rohit Ghai, Oded Beja, Hideki Kandori
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Journal Title
Science Advances
Volume: 6
Issue: 15
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research
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[Presentation] 第58回日本生物物理学会年会2020
Author(s)
長坂 勇次郎、細島 頌子、神取 秀樹、井上 圭一、八尾 寛
Organizer
The gate-keeper role of a highly conserved helix-3 tryptophan for ion transport of the channelrhodopsin chimera, C1C2/ChRWR
Related Report
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