2021 Fiscal Year Research-status Report
Competitive linear ubiquitin binding in NFkB-signaling
| Project/Area Number |
20K15756
|
|---|
| Research Institution | Kyoto University |
|---|
Principal Investigator |
Walinda Erik 京都大学, 医学研究科, 助教 (80782391)
|
|---|
| Project Period (FY) |
2020-04-01 – 2023-03-31
|
| Keywords | ubiquitin / polyubiquitin / binding domains / Protein binding / Molecular competition / Linear polyubiquitin / NF-kappa B |
|---|
| Outline of Annual Research Achievements |
Ubiquitin chains and binding proteins have been successfully purified with great yield, purity, and reproducibility. ITC and NMR experiments proceeded smoothly as anticipated and future studies are on the way. Both all-atom and coarse grained MD simulations under the MARTINI force field also indicate interesting molecular behavior and hint at intriguing mechanisms for the physiological implications of the competition mechanism.
Due to the pandemic situation, the current data could not yet be presented at any international conferences as had been originally scheduled. Instead, a publication of the NMR and MD data of solution state NMR binding to linear ubiquitin chains including structural data is in preparation. Conference presentation of the current data and conclusions to be drawn from this project is currently delayed until August (ICMRBS 2022).
|
| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
Thanks to acquisition of valuable experimental equipment and ample experimental time allocation on the spectrometer, the necessary basic NMR tasks such as signal assignment have been completed. Titration data have also been gathered and analyzed. Competition experiments using both NMR and ITC have been carried out.
|
| Strategy for Future Research Activity |
Construction and validation of theoretical models that accurately reflect the molecular competition between A20, NEMO, and HOIL-1L for binding to linear ubiquitin chains is warranted.
|
| Causes of Carryover |
As pointed out in the research status report, several experiments are yet to be conducted. Moreover, publication of the results depends on these experiments; therefore publication fees have to be accounted for in the next fiscal year.
|
