Protein 19F Labeling Strategy for In-Cell NMR and Functional Analyses

2017 Fiscal Year Final Research Report

• Project/Area Number: 26440018
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: Tohoku University
• Principal Investigator: Koshiba Seizo 東北大学, 東北メディカル・メガバンク機構, 教授 (70332301)
• Project Period (FY): 2014-04-01 – 2018-03-31
• Keywords: フッ素 / タンパク質 / NMR / 細胞内環境 / 相互作用 / 動的構造変化

Outline of Final Research Achievements

Fluorine 19 (19F) is a widely used nucleus for NMR because of its high sensitivity and large chemical shift dispersion. In particular, 19F-labeling is useful for In-Cell NMR, because most of biological systems contain no fluorine, resulting that 19F-labeled molecules in living cells can be selectively analyzed. In this study, I developed several 19F-labeling methods for proteins and effectively incorporated trifluoromethyl-labeled amino acids into proteins. I expressed several kinds of trifluoromethyl-labeled proteins and could observe highly sensitive 19F NMR signals both in vitro and in vivo. I also investigated the interactions of ligands with large molecular weight proteins by 19F-NMR and could identify binding sites of the ligands in a short time. These results show that my methods are particularly useful for investigating behavior of intracellular proteins or ligand-interactions. I expect that this labeling strategy is useful for wide variety of biological sciences.

Free Research Field

NMRによるタンパク質の構造生物学