Reconstitution of private chaperone systems using synthetic binding proteins

2018 Fiscal Year Final Research Report

• Project/Area Number: 15H05370
• Research Category: Grant-in-Aid for Young Scientists (A)
• Allocation Type: Single-year Grants
• Research Field: Biomolecular chemistry
• Research Institution: Okayama University
• Principal Investigator: Yasui Norihisa 岡山大学, 医歯薬学総合研究科, 准教授 (90467514)
• Project Period (FY): 2015-04-01 – 2019-03-31
• Keywords: 人工タンパク質 / タンパク質 / 進化分子工学 / 合成生物学 / 分子シャペロン / 一本鎖モネリン

Outline of Final Research Achievements

In this study, to clarify the correlation between the fold-specific substrate recognition and the function of the private chaperones, the system for generation of the synthetic binding proteins that specifically bind to the target proteins has been developed. The phage display libraries of the synthetic binding proteins using single-chain monellin as a non-antibody molecular scaffold were designed and constructed. By sorting the phage display libraries against the green fluorescent protein variant (GFPuv), the synthetic binding protein that has the ability to bind to GFPuv (GBP-1) was generated. The characteristics of GBP-1 revealed in this study indicate that single-chain monellin is useful as a non-antibody molecule backbone of the synthetic binding proteins.

Free Research Field

タンパク質工学

Academic Significance and Societal Importance of the Research Achievements

本研究では，ファージディスプレイライブラリーの作製とその選別により検討を重ねた結果，一本鎖モネリンを非抗体分子骨格として利用すると，標的分子に親和性を示す人工結合タンパク質を作製できることを示した。本研究で構築した一本鎖モネリンを非抗体分子骨格とする人工結合タンパク質ライブラリーは，プライベートシャペロンの機能解析に留まらず，様々な研究で利用することができる点でも意義がある。