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2021 年度 実施状況報告書

NF-κB経路における直鎖状ユビキチン鎖の競合的結合

研究課題

研究課題/領域番号 20K15756
研究機関京都大学

研究代表者

Walinda Erik  京都大学, 医学研究科, 助教 (80782391)

研究期間 (年度) 2020-04-01 – 2023-03-31
キーワードubiquitin / polyubiquitin / binding domains / Protein binding / Molecular competition / Linear polyubiquitin / NF-kappa B
研究実績の概要

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).

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

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.

今後の研究の推進方策

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.

次年度使用額が生じた理由

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.

  • 研究成果

    (6件)

すべて 2022 2021

すべて 雑誌論文 (6件)

  • [雑誌論文] Structural Insights into Methylated DNA Recognition by the Methyl-CpG Binding Domain of MBD6 from <i>Arabidopsis thaliana</i>2022

    • 著者名/発表者名
      Mahana Yutaka、Ohki Izuru、Walinda Erik、Morimoto Daichi、Sugase Kenji、Shirakawa Masahiro
    • 雑誌名

      ACS Omega

      巻: 7 ページ: 3212~3221

    • DOI

      10.1021/acsomega.1c04917

  • [雑誌論文] Multiple-State Monitoring of SOD1 Amyloid Formation at Single-Residue Resolution by Rheo-NMR Spectroscopy2021

    • 著者名/発表者名
      Iwakawa Naoto、Morimoto Daichi、Walinda Erik、Shirakawa Masahiro、Sugase Kenji
    • 雑誌名

      Journal of the American Chemical Society

      巻: 143 ページ: 10604~10613

    • DOI

      10.1021/jacs.1c02974

  • [雑誌論文] Backbone resonance assignments of the A2 domain of mouse von Willebrand factor2021

    • 著者名/発表者名
      Morimoto Daichi、Osugi Masanori、Mahana Yutaka、Walinda Erik、Shirakawa Masahiro、Sugase Kenji
    • 雑誌名

      Biomolecular NMR Assignments

      巻: 15 ページ: 427~431

    • DOI

      10.1007/s12104-021-10041-8

  • [雑誌論文] Effects of Weak Nonspecific Interactions with ATP on Proteins2021

    • 著者名/発表者名
      Nishizawa Mayu、Walinda Erik、Morimoto Daichi、Kohn Benjamin、Scheler Ulrich、Shirakawa Masahiro、Sugase Kenji
    • 雑誌名

      Journal of the American Chemical Society

      巻: 143 ページ: 11982~11993

    • DOI

      10.1021/jacs.0c13118

  • [雑誌論文] Expression, solubility monitoring, and purification of the co-folded LUBAC LTM domain by structure-guided tandem folding in autoinducing cultures2021

    • 著者名/発表者名
      Walinda Erik、Morimoto Daichi、Sorada Tomoki、Iwai Kazuhiro、Sugase Kenji
    • 雑誌名

      Protein Expression and Purification

      巻: 187 ページ: 105953~105953

    • DOI

      10.1016/j.pep.2021.105953

  • [雑誌論文] Rigorous analysis of the interaction between proteins and low water-solubility drugs by qNMR-aided NMR titration experiments2021

    • 著者名/発表者名
      Hirakawa Takuya、Walinda Erik、Morimoto Daichi、Sugase Kenji
    • 雑誌名

      Physical Chemistry Chemical Physics

      巻: 23 ページ: 21484~21488

    • DOI

      10.1039/D1CP03175A

URL: 

公開日: 2023-12-25  

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