In-situ structural biology by NMR spectroscopy

2023 Fiscal Year Final Research Report

• Project/Area Number: 21H02419
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 43020:Structural biochemistry-related
• Research Institution: Tokyo Metropolitan University
• Principal Investigator: Ito Yutaka 東京都立大学, 理学研究科, 教授 (80261147)
• Co-Investigator(Kenkyū-buntansha): 美川 務 国立研究開発法人理化学研究所, 生命機能科学研究センター, 専任研究員 (20321820)
• Project Period (FY): 2021-04-01 – 2024-03-31
• Keywords: In-cell NMR / 蛋白質 / 立体構造 / ダイナミクス / 相互作用 / 液液相分離

Outline of Final Research Achievements

We have studied on (1) analysis of three-dimensional structures and dynamics of proteins in live cells, (2) analysis of inter-domain interactions of multi-domain proteins in live cells, and (3) analysis of intracellular protein-protein interactions.
For (1), we developed Key elemental technologies such as "methods for rapid multidimensional NMR measurements and efficient reconstruction of spectra", "automated NMR data analysis and high-precision 3D structure determination of proteins using information science" and "Bioreactor microculture techniques for long-term NMR measurement of cell samples". For (2), the relative domain orientations were determined for proteins such as linear-polyUb by using PCS/PRE paramagnetic NMR data. For (3), results were obtained in the analysis of the interaction between Nrf2 and Keap1, etc. in human cells.

Free Research Field

構造生物化学

Academic Significance and Societal Importance of the Research Achievements

本研究の成果によって，これまで困難であった，「細胞内環境が，様々な生命現象に関与する蛋白質群の立体構造やダイナミクスに与える影響」の詳細な解析が，in-cell NMRの手法を用いて可能になりつつある．細胞内クラウディング環境は生体高分子が内在的に持つ生物活性上重要な性質を顕在化させている可能性があり，単離精製された状態に加えて，細胞内での蛋白質動態の研究が加速すれば，蛋白質の生物活性発現のより良い理解につながる．また，本研究の成果は，例えばヒト培養細胞内の原子分解能でのドラッグスクリーニングなど，医療や創薬への波及効果も大いに期待される．