Development of reverse-protein splicing method using proteases
| Project/Area Number |
18K05113
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Review Section |
Basic Section 33020:Synthetic organic chemistry-related
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| Research Institution | Tokai University |
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Principal Investigator |
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| Project Period (FY) |
2018-04-01 – 2021-03-31
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| Project Status |
Completed (Fiscal Year 2020)
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| Budget Amount *help |
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2020: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2019: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2018: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
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| Keywords | タンパク質逆スプライシング / プロテアーゼ / ネイティブケミカルライゲーション / タンパク質逆スプライシング反応 / ペプチド縮合反応 / Native chemical ligation / ペプチドライゲーション / プロテーゼ |
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| Outline of Final Research Achievements |
Protein splicing is found in nature, although its reverse reaction has not yet been developed. In this project, we tried to develop reverse-protein splicing (RPS) reaction using a protease as catalyst. When using lysyl endopeptidase (LEP) as the catalyst, RPS reaction proceeded in ethylene glycol aqueous solution, but the yield of desired product was low. The peptide with non-proteinogenic amino acid was also acted as an insert in this reaction. This is the first case for one-pot chemical RPS reaction, and it might be useful for the regioselective chemical modification of polypeptides.
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| Academic Significance and Societal Importance of the Research Achievements |
有機化学的にRPS反応を行った例は、これまでに報告されていないことから、世界初の実施例であると考えられる。この方法を利用すれば、将来的に組換えタンパク質の部位特異的なアミノ酸配列変換などに利用できるものと期待される。また、非タンパク質性の化学構造も挿入可能であったことから、蛍光団の挿入などの部位特異的な化学修飾にも応用可能であり、新たなタンパク質化学修飾法として生化学分野、ケミカルバイオロジー分野への応用が期待される。
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Report
(4 results)
Research Products
(9 results)
