Detection system of the intracellular nitric oxide in yeast by HPLC with a fluorescence detector.

Nitric oxide (NO) is an important signaling molecule involved in various biological phenomena in many organisms. The physiological functions and metabolism of NO in yeast, a unicellular microorganism, are still unknown, mainly because it is difficult to analyze the intracellular NO levels accurately. Here, we developed a new method of more accurately measuring NO content in yeast cells with the detection limit of 6 nM, by treating the cells with an NO-specific fluorescence probe followed by high-performance liquid chromatography with fluorescence detection (HPLC/FLD). This approach successfully detected and quantified the NO content inside yeast cells treated with an NO donor. Moreover, the HPLC/FLD analysis indicates that the fluorescence induced under some environmental stress conditions, such as ethanol, vanillin, and heat-shock, was not derived from NO. The HPLC/FLD method developed in this study provides a new strategy for measuring the intracellular NO concentration with higher accuracy. Copyright © 2020 Elsevier Inc. All rights reserved.

Citation

Ryo Nasuno, Seiya Shino, Yuki Yoshikawa, Natsuko Yoshioka, Yuichi Sato, Kohei Kamiya, Hiroshi Takagi. Detection system of the intracellular nitric oxide in yeast by HPLC with a fluorescence detector. Analytical biochemistry. 2020 Jun 01;598:113707

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PMID: 32251648

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