Establishment of a new protein structure stabilization strategy based on synthetic binding proteins

2023 Fiscal Year Final Research Report

• Project/Area Number: 21K05386
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 38030:Applied biochemistry-related
• Research Institution: Kyoto Prefectural University
• Principal Investigator: Tanaka Shun-ichi 京都府立大学, 生命環境科学研究科, 准教授 (70591387)
• Co-Investigator(Kenkyū-buntansha): 松村 浩由 立命館大学, 生命科学部, 教授 (30324809) ; 高野 和文 京都府立大学, 生命環境科学研究科, 教授 (40346185) ; 吉澤 拓也 立命館大学, 生命科学部, 講師 (50779056)
• Project Period (FY): 2021-04-01 – 2024-03-31
• Keywords: タンパク質工学 / 酵素 / 人工結合タンパク質 / 安定性 / 熱力学解析 / 立体構造解析

Outline of Final Research Achievements

In this study, we aimed to elucidate the general applicability of the protein's structure stabilization effect induced by the binding of synthetic binding proteins and to understand the mechanism of how binding to specific sites and in specific modes leads to stabilization. To achieve this, we conducted comprehensive and systematic analyses using thermodynamic and structural methods. We successfully generated synthetic binding proteins for various types of targets and performed affinity analysis using surface plasmon resonance (SPR) as well as structural analysis using X-ray crystallography, X-ray small-angle scattering, and cryo-electron microscopy. However, progress in thermodynamic analysis has been slow, and we have not yet drawn conclusions about the correlation with stabilization. Moving forward, we will continue our research with the aim of achieving the initial goals by advancing the thermodynamic analysis.

Free Research Field

応用生物化学関連

Academic Significance and Societal Importance of the Research Achievements

今後継続する研究成果次第ではあるが、人工結合タンパク質の相互作用部位・様式と安定化効果との相関関係を詳細に明らかにすることができれば、変異導入に依存してきた従来の方法論とは異なる新たなタンパク質安定化戦略の創成に繋がるため、その学術的意義は大きいと考えられる。また、この新たなタンパク質安定化戦略によって、従来の手法では難しく諦められていた酵素の改変が広く可能となれば、様々な新しい食品素材、医薬品、工業原料などの創出に繋がるため、産業的意義も大きいと考えられる。