Visualization of NO3−/NO2− Dynamics in Living Cells by Fluorescence Resonance Energy Transfer (FRET) Imaging Employing a Rhizobial Two-component Regulatory System

• DOI: 10.1074/jbc.m115.687632
• Publication Date: 2015-12-03T01:40:03Z
• Authors: Masafumi Hidaka; Aina Gotoh; Taiki Shimizu; Kiwamu Minamisawa; Hiromi Imamura; Takafumi Uchida
• Keywords: Two-component regulatory system

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