Structural analysis of protein intrinsically disordered regions that drive phase transition

2023 Fiscal Year Final Research Report

• Project/Area Number: 19K14677
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 13040:Biophysics, chemical physics and soft matter physics-related
• Research Institution: Osaka University
• Principal Investigator: Yoshimura Yuichi 大阪大学, 蛋白質研究所, 招へい研究員 (70632248)
• Project Period (FY): 2019-04-01 – 2024-03-31
• Keywords: 天然変性蛋白質 / 核磁気共鳴 / カルボニル１３Ｃ

Outline of Final Research Achievements

This study was focused on structural dynamics of protein intrinsically disordered regions that drive phase transition. To this end, nuclear magnetic resonance (NMR) experiments were presented that utilize carbonyl 13C direct-detection. A 13C-detect 2D (HACA)CON spectrum provided well-dispersed resonances, and these resonances circumvented solvent exchange broadening. Sequential resonance assignments were made through the 13C-detect 3D (HA)CACON and (HA)CANCO experiments. A method relying on the detection of the anti-phase operator 2NxHz decorrelation was used to obtain the chemical shifts and solvent exchange rates of individual amide protons in the protein. The approach demonstrated in this study may be widely used to analyze structural dynamics of intrinsically disordered proteins.

Free Research Field

生物物理学

Academic Significance and Societal Importance of the Research Achievements

生体内での機能上の役割が注目される蛋白質の天然変性領域は、動的に揺動するなかで機能制御を実現する。柔らかな構造を有する変性領域は蛋白質の結晶化を妨げるため、従来の結晶構造解析の適用は困難である。一方で、蛋白質分子を原子分解能で観測する核磁気共鳴（ＮＭＲ）は、溶液中での構造動態解析が可能であり、相互作用解析や夾雑環境での解析において威力を発揮する。本研究で開発した新規ＮＭＲ測定法を適用することで、蛋白質の天然変性領域が駆動する液液相分離（ＬＬＰＳ）の分子機構の解明やＬＬＰＳを標的とした創薬研究への展開が期待される。