Journal
REDOX BIOLOGY
Volume 18, Issue -, Pages 295-304Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.redox.2018.07.016
Keywords
Polysulfide quantification; LC-MS/MS; SSP4; Biological samples
Categories
Funding
- European Society of Cardiology [R-2016-054]
- Schering Stiftung
- Deutsche Forschungsgemeinschaft [SFB 815/A16, SFB 834/B13, Exzellenzcluster 147]
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Hydrogen sulfide (H2S) is a gasotransmitter that regulates cellular homeostasis and impacts on multiple physiological and pathophysiological processes. However, it exerts many of its biological actions indirectly via the formation of H2S-derived sulfane sulfur species/polysulfides. Because of the high reactivity of sulfur species, the detection of H2S-derived polysulfides in biological systems is challenging and currently used methods are neither sensitive nor quantitative. Herein, we describe a LC-MS/MS-based method that makes use of Sulfane Sulfur Probe 4 to detect endogenously generated polysulfides in biological samples in a selective, sensitive and quantitative manner. The results indicate a large variability in the activity of the H2S-generating enzymes in different murine organs, but the method described was able to detect intracellular levels of polysulfides in the nanomolar range and identify cystathionine gamma-lyase as the major intracellular source of sulfane sulfur species/ polysulfides in murine endothelial cells and hearts. The protocol described can be applied to a variety of biological samples for the quantification of the H2S-derived polysulfides and has the potential to increase understanding on the control and consequences of this gaseous transmitter.
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