In-cell NMR studies of metalloproteins

2023 Fiscal Year Final Research Report

• Project Area: Integrated Biometal Science: Research to Explore Dynamics of Metals in Cellular System
• Project/Area Number: 19H05773
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Complex systems
• Research Institution: Tokyo Metropolitan University
• Principal Investigator: Ito Yutaka 東京都立大学, 理学研究科, 教授 (80261147)
• Project Period (FY): 2019-06-28 – 2024-03-31
• Keywords: In-cell NMR / 溶液NMR / 金属結合蛋白質 / 立体構造

Outline of Final Research Achievements

We have studied on (1) development of in-cell NMR methods, (2) analysis of biometal-related proteins using solution and in-cell NMR spectroscopy, and (3) potential applications of in-cell NMR.
In (1), we developed an in-cell NMR method using cultured plant cells and a method for 3D structural analysis of multi-domain proteins using paramagnetic NMR information. In (2), we analysed the rhizobial two-component signal transduction system FixJ/FixL system. For FixJ, we determined the detailed solution structures as well as analysing the interaction with FixL and the effect of phosphorylation. In (3), results were obtained for in-cell NMR analysis of unstable proteins in human cultured cells.

Free Research Field

構造生物化学

Academic Significance and Societal Importance of the Research Achievements

In-cellおよび溶液NMR測定・解析技術の高度化と汎用化を行うことで，これまで解析が困難であった様々な金属結合蛋白質の溶液状態の動態，細胞内の動態を詳細に解析することができるようになった．細胞内でのマルチドメイン蛋白質のドメイン間相対配置の解析も可能になりつつある．特にFixJの溶液構造解析や，FixLとの相互作用やリン酸化の解析では生物学的に興味深い結果を得ることができた．In-cell NMRは，細胞内の標的蛋白質と低分子の相互作用を指標にした阻害剤探索にも応用可能である．基礎科学のみならず，先端医療や創薬科学などの産業にも波及的効果が期待できる．