2021 Fiscal Year Final Research Report
Application of R1EN-Fusion method to large proteins
| Project/Area Number |
19K05696
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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 37010:Bio-related chemistry
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| Research Institution | National Institutes for Quantum Science and Technology (2020-2021)
The University of Tokushima (2019) |
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Principal Investigator |
MAITA NOBUO 国立研究開発法人量子科学技術研究開発機構, 量子生命科学研究所, 上席研究員 (00404046)
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| Project Period (FY) |
2019-04-01 – 2022-03-31
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| Keywords | 構造生物学 / Ｘ線結晶構造解析 |
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| Outline of Final Research Achievements |
The R1EN-fusion method is a simple way to determine the crystal structure of fused-guest proteins. However, the upper limit of molecular weight is about 21 kDa, which may be a bottleneck for versatility. To overcome this issue, I constructed the disulfide-bonded dimer and trimer of R1EN and tried to crystallize them, but the success rate of crystallization was poor. Thus, I searched for the crystallization conditions again using Tris-buffer and obtained a thin, elongated crystals at pH 7.4. I expect that it will be able to refine the conditions and obtain crystals more consistently and applicable to structural study of large proteins.
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| Free Research Field |
構造生物化学
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| Academic Significance and Societal Importance of the Research Achievements |
タンパク質の結晶構造解析は、基礎生物学から疾病、薬剤開発にわたって有用な知見を与える。しかし結晶化は不確定要素が多くハードルが高い。本研究は、簡単にタンパク質結晶を得ることが出来るR1EN-fusion法を、より広範囲の大きさのタンパク質に適用するための研究である。これによりこれまで構造不明であったタンパク質の詳細な立体構造が解明できると期待される。
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