TY - JOUR
T1 - Visualization of NO3-/NO2- dynamics in living cells by fluorescence resonance energy transfer (FRET) imaging employing a rhizobial two-component regulatory system
AU - Hidaka, Masafumi
AU - Gotoh, Aina
AU - Shimizu, Taiki
AU - Minamisawa, Kiwamu
AU - Imamura, Hiromi
AU - Uchida, Takafumi
N1 - Publisher Copyright:
© 2016 by The American Society for Biochemistry and Molecular Biology, Inc.
PY - 2016/1/29
Y1 - 2016/1/29
N2 - 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.
AB - 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.
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U2 - 10.1074/jbc.M115.687632
DO - 10.1074/jbc.M115.687632
M3 - Article
C2 - 26631727
AN - SCOPUS:84956982850
SN - 0021-9258
VL - 291
SP - 2260
EP - 2269
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 5
ER -