Visualization of NO3-/NO2- dynamics in living cells by fluorescence resonance energy transfer (FRET) imaging employing a rhizobial two-component regulatory system

Masafumi Hidaka, Aina Gotoh, Taiki Shimizu, Kiwamu Minamisawa, Hiromi Imamura, Takafumi Uchida

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Nitrate (NO3-) and nitrite (NO2-) are the physiological sources of nitric oxide (NO), a key biological messenger molecule.NO3-/NO2- exerts a beneficial impact on NO homeostasis and its related cardiovascular functions. To visualize the physiological dynamics of NO3-/NO2- for assessing the precise roles of these anions, we developed a genetically encoded intermolecular fluorescence resonance energy transfer (FRET)-based indicator, named sNOOOpy (sensor for NO3-/NO2- in physiology), by employing NO3-/NO2--induced dissociation of NasST involved in the denitrification system of rhizobia. The in vitro use of sNOOOpy shows high specificity for NO3- and NO2-, and its FRET signal is changed in response to NO3-/NO2- in the micromolar range. Furthermore, both an increase and decrease in cellular NO3- concentration can be detected. sNOOOpy is very simple and potentially applicable to a wide variety of living cells and is expected to provide insights into NO3-/NO2- dynamics in various organisms, including plants and animals.

Original languageEnglish
Pages (from-to)2260-2269
Number of pages10
JournalJournal of Biological Chemistry
Volume291
Issue number5
DOIs
Publication statusPublished - 2016 Jan 29

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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