Τίτλος:
A selective and sensitive method for quantification of endogenous polysulfide production in biological samples
Γλώσσες Τεκμηρίου:
Αγγλικά
Περίληψη:
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 γ-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. © 2018 The Authors
Συγγραφείς:
Bibli, S.-I.
Luck, B.
Zukunft, S.
Wittig, J.
Chen, W.
Xian, M.
Papapetropoulos, A.
Hu, J.
Fleming, I.
Λέξεις-κλειδιά:
cystathionine gamma lyase; fluorescent dye; hydrogen sulfide; polysulfide; sulfide; unclassified drug; cystathionine gamma lyase; polysulfide; sulfide, animal cell; animal experiment; animal tissue; Article; biological activity; biological model; complex formation; controlled study; endothelium cell; enzyme activity; female; heart; human; human cell; limit of detection; liquid chromatography-mass spectrometry; male; mouse; nonhuman; priority journal; regulatory mechanism; animal; C57BL mouse; cardiac muscle; cell culture; chemistry; enzymology; liquid chromatography; metabolism; procedures; tandem mass spectrometry, Animals; Cells, Cultured; Chromatography, Liquid; Cystathionine gamma-Lyase; Endothelial Cells; Humans; Mice, Inbred C57BL; Myocardium; Sulfides; Tandem Mass Spectrometry
DOI:
10.1016/j.redox.2018.07.016
