Ensemble structure determination using multiple measurement data in solution state

2023 Fiscal Year Final Research Report

• Project/Area Number: 21K06114
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 43040:Biophysics-related
• Research Institution: Tokyo Metropolitan University
• Principal Investigator: Ikeya Teppei 東京都立大学, 理学研究科, 准教授 (30457840)
• Project Period (FY): 2021-04-01 – 2024-03-31
• Keywords: GRB2 / SOS1 / 多価相互作用 / 液液相分離 / アンサンブル構造解析 / 天然変性タンパク質 / マルチドメインタンパク質

Outline of Final Research Achievements

We developed two new programs to calculate intermolecular binding affinity from NMR interaction analysis and applied them to analyze the interactions between GRB2-SOS1 and Drk-Sos. As a result, it was revealed that the N-terminal SH3 domains of GRB2 and Drk bind more strongly to SOS than the C-terminal SH3 domains. Based on this, we proposed a molecular recognition model for SOS by GRB2 and Drk, providing new insights into the LLPS formation mechanism by GRB2-SOS1. Additionally, using the multi-state structure calculation method, we analyzed the solution structure of GRB2 and clarified that two of its domains are united while one remains independently mobile. Furthermore, we analyzed LLPS formation by GRB2 and SOS1 using solution NMR.

Free Research Field

生物物理

Academic Significance and Societal Importance of the Research Achievements

液液相分離（LLPS）は，多くの細胞内イベントで重要な役割を果たしている．LLPSは常に動的な変化を伴い，マルチドメイン蛋白質，天然変性蛋白質，RNAなどの柔軟性の高い分子が関与している．溶液NMRは，こうした分子の動的変化を原子分解能で解析できる有力な手法である．本研究で開発した計算手法やNMR計測法を応用することで，LLPS形成機構に関する構造生物学的知見がさらに深まることが期待される．