Elucidation of effect of phase separation on conformation and dynamics of spider silk proteins
| Project/Area Number |
20K15738
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| Research Category |
Grant-in-Aid for Early-Career Scientists
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| Allocation Type | Multi-year Fund |
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| Review Section |
Basic Section 43020:Structural biochemistry-related
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| Research Institution | Institute of Physical and Chemical Research |
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Principal Investigator |
Oktaviani Nur Alia 国立研究開発法人理化学研究所, 環境資源科学研究センター, 基礎科学特別研究員 (70753838)
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| Project Period (FY) |
2020-04-01 – 2023-03-31
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| Project Status |
Completed (Fiscal Year 2022)
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| Budget Amount *help |
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2022: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2021: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2020: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
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| Keywords | NMR / spider silk / protein / structure / mechanism / LLPS / spider silk protein / タンパク質NMR / タンパク質の構造とダイナミクス / 生体高分子 / dynamics |
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| Outline of Research at the Start |
Upon biomimetic spinning process, liquidliquid
phase separation of spidroin results in the formation of a high-density phase, which is separated from the low-density phase for further processing. Understanding the mechanism of biomimetic spinning is very essential for development of biomimetic spinning of artificial spidroin, which is still poorly understood until now. In this proposed research, the conformation and dynamics during liquid-liquid phase separation of spidroin will be elucidated using multidimensional NMR spectroscopy combined with other biophysical tools.
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| Outline of Final Research Achievements |
In this fiscal year, I investigated the effect of phase separation on conformation and dynamics of the repetitive domain and full length minispidroin NR6C and NR12C. The tyrosine residue of the repetitive domain was found to be affected upon LLPS phenomenon. Then, we made a mutant on the repetitive domain which substituted the tyrosine with serine residue. The mutant does not have an effect on the conformattion and dynamics of minispidroin. However, we found a drastic effect on the mutant, where the oligomerization was suppressed at higher kPi concentration, suggesting the important role of tyrosine residues in promoting oligomerization upon LLPS phenomenon. Overall, we found that C-terminal domain (CTD) and tyrosine from the repetitive domain are important as stickers which mediate the oligomerization on the LLPS formation.
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| Academic Significance and Societal Importance of the Research Achievements |
In natural spinning process, spidroins undergo the liquid-liquid phase separation (LLPS). Since LLPS is one of the emerging topics in biology, thus this study is useful not only to understand the mechanism of natural spinning process, but also to obtain the insight into the molecular basis of LLPS.
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Report
(3 results)
Research Products
(8 results)
