2017年2月17日
Development of an Azoreductase-based Reporter System with Synthetic Fluorogenic Substrates
ACS Chemical Biology
- 巻
- 12
- 号
- 2
- 開始ページ
- 558
- 終了ページ
- 563
- 記述言語
- 英語
- 掲載種別
- 研究論文(学術雑誌)
- DOI
- 10.1021/acschembio.6b00852
- 出版者・発行元
- AMER CHEMICAL SOC
Enzyme/substrate pairs, such as beta-galactosidase with chromogenic x-gal substrate, are widely used as reporters to monitor biological events, but there is still a requirement for new reporter systems, which may be orthogonal to existing systems. Here, we focused on azoreductase (AzoR). We designed and synthesized a library of azo-rhodamine derivatives as candidate fluorogenic substrates. These derivatives were nonfluorescent, probably due to ultrafast conformational change around the N=N bond after photoexcitation. We found that AzoR-mediated reduction of the azo bond of derivatives bearing an electron donating group on the azobenzene moiety was followed by nonenzymatic cleavage to afford highly fluorescent 2-methyl-rhodamine green (2-Me RG), which was well retained in cells. We show that the AzoR/compound 9 reporter system can detect azoreductase-expressing live cells at the single cell level.
- リンク情報
- ID情報
-
- DOI : 10.1021/acschembio.6b00852
- ISSN : 1554-8929
- eISSN : 1554-8937
- PubMed ID : 28036168
- Web of Science ID : WOS:000394736700030