| Project/Area Number |
20K15756
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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 43040:Biophysics-related
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| Research Institution | Kyoto University |
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Principal Investigator |
WALINDA Erik 京都大学, 医学研究科, 助教 (80782391)
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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 |
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2021: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2020: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
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| Keywords | ubiquitin / ubiquitin binding / NZF domain / M1 ubiquitin chains / linear ubiquitin chains / NMR spectroscopy / ITC / MD simulations / M1-linked chains / polyubiquitin / binding domains / Protein binding / Molecular competition / Linear polyubiquitin / NF-kappa B / Ubiquitin |
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| Outline of Research at the Start |
[1] Binding of NEMO to linear ubiquitin in vitro, [2] Binding of A20 to linear ubiquitin in vitro
First we will conduct the binary experiments for both proteins and other similar candidates such as HOIL-1L binding to linear ubiquitin in vitro. These parameters will be necessary to dissect the competitive mechanism [3]
[3] Competitive binding to linear ubiquitin in vitro
ITC and NMR experiments coupled with atomic simulations together will provide a comprehensive picture of the competition between A20 and NEMO (and HOIL-1L) for binding to linear ubiquitin.
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| Outline of Final Research Achievements |
In these two yearsl our research has proceeded rather smoothly. We were able to publish several aspects of this project in various international peer-reviewed journals.
The project focuses on protein ubiquitylation, attaching ubiquitin to target proteins. Initially, it studied K48-linked polyubiquitin chains for degradation. The discovery of linear ubiquitin chains and LUBAC's role in immune signaling necessitated understanding them further. The study aims to explore HOIL-1L's binding specificity to linear ubiquitin chains, conformational changes, preference for M1-linked polyubiquitin, and its role in HOIL-1L and LUBAC. Key findings include optimized purification of LUBAC fragments for cancer drug screening, investigating cyclization's impact on ubiquitin chain recognition by OTUB1, and examining linear ubiquitin recognition by HOIL-1L NZF domain. These results enhance understanding of linear ubiquitin binding and the ubiquitin code.
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| Academic Significance and Societal Importance of the Research Achievements |
Scientific significance: understanding non-proteolytic functions of ubiquitin chains, particularly in immune signaling, survival, inflammation, immune disorders, cancer, and infectious diseases.
Social significance: potential to contribute to the development of treatments for immune disorders.
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