Turn-on fluorescent labeling of target protein at a specific lysine residue and formation of protein cross-link with fluorescence for therir application to protein studies

2022 Fiscal Year Final Research Report

• Project/Area Number: 18K05319
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 37010:Bio-related chemistry
• Research Institution: Kyushu University
• Principal Investigator: Aso Mariko 九州大学, 薬学研究院, 准教授 (30201891)
• Project Period (FY): 2018-04-01 – 2023-03-31
• Keywords: リジン修飾 / 環境応答型蛍光 / 反応性核酸

Outline of Final Research Achievements

We investigated new molecules which convert lysine residue in target protein into environment sensitive fluorophore and provide fluorescent-labeled proteins to monitor protein interaction. Fluorescent labeling of nucleic acid binding protein proceeded with the molecule carrying nucleic acid. Difference in fluorescence intensities was observed between modified proteins in the free and DNA bound forms. Introduction of substituents into fluorophore provided new molecules and fluorophore more stable than the original compound was obtained. We also studied molecules which convert lysine into fluorophore with other functions.

Free Research Field

生体分子関連化学

Academic Significance and Societal Importance of the Research Achievements

今回開発したリジン修飾分子は蛋白質親和性の核酸部をもち、標的蛋白質に結合し、緩和な条件でリジン残基を環境応答型蛍光基に変換することができた。修飾蛋白質は核酸の結合の有無により、蛍光強度の変化がみられることが分かり、蛋白質相互作用の検出する分子設計の妥当性を示した。また安定性が向上した蛍光分子や異なる物性を持つ蛍光分子を見出し、これらを生成する修飾分子の合成法を確立した。これらの成果により本手法を用いた蛋白質修飾の更なる展開が可能となった。