| 研究課題/領域番号 |
22K06161
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| 研究種目 |
基盤研究(C)
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| 配分区分 | 基金 |
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| 応募区分 | 一般 |
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| 審査区分 |
小区分43040:生物物理学関連
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| 研究機関 | 京都大学 |
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研究代表者 |
Walinda Erik 京都大学, 医学研究科, 助教 (80782391)
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| 研究期間 (年度) |
2022-04-01 – 2025-03-31
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| 研究課題ステータス |
交付 (2023年度)
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| 配分額 *注記 |
4,160千円 (直接経費: 3,200千円、間接経費: 960千円)
2024年度: 650千円 (直接経費: 500千円、間接経費: 150千円)
2023年度: 1,300千円 (直接経費: 1,000千円、間接経費: 300千円)
2022年度: 2,210千円 (直接経費: 1,700千円、間接経費: 510千円)
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| キーワード | ubiquitin / linear ubiquitin / ubiquitin binding / M1-linked chains / HOIL-1L / NZF domain / zinc finger / Ubiquitin / NF-kB signaling / Binding proteins / Chemical physics / Competitive binding |
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| 研究開始時の研究の概要 |
The outline is as follows:
[1] Analyze HOIL-1L, NEMO, A20 binding to linear ubiquitin in vitro.
First we will conduct the binary experiments for these both proteins for various chain lengths of linear polyubiquitin. These parameters will be necessary to dissect the competitive mechanism.
[2] Competitive binding to linear ubiquitin in vitro.
We will couple ITC, fluorescence spectroscopy, and NMR experiments together with molecular simulations to obtain a comprehensive picture of the competition between A20, NEMO, and HOIL-1L for binding to linear ubiquitin to aid NFkB fundamental understanding.
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| 研究実績の概要 |
Focusing on the linear ubiquitin chain assembly complex, we elucidate how it synthesizes "head-to-tail" poly-Ub chains crucial for immune signaling and cell death regulation. Specifically, we investigate the interaction between HOIL-1L and linear poly-Ub chains, revealing the molecular determinants driving their selective binding. Through NMR and biophysical methods, we unveil the dynamic process by which the NZF domain of HOIL-1L evolves into the specific linear di-Ub-bound state while excluding other potential Ub species. Our findings highlight the role of conserved electrostatic contacts and the impact of phosphorylation at threonine-207 on linear Ub affinity. This research deepens our understanding of the Ub code and offers insights valuable for immune diseases and cancer research.
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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
2: おおむね順調に進展している
理由
Overall, the research is progressing smoothly. However, further analysis and validation experiments are needed. While we are making large progress on understanding the kinetics of binary systems, consisiting of ubiquitin chains and their rececptors, it is tricky to dissect the competitve systems. However, we have experiments lined up that can surely elucidate this elusive mechanism.
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| 今後の研究の推進方策 |
We will continue to dissect the kinetics of ubiquitin recognition by HOIL-1L and similar ubiquitin-binding proteins by using biophysical methods.
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