Recent developments in detection of superoxide radical anion and hydrogen
peroxide: Opportunities, challenges, and implications in redox signaling
#MMPMID27590268
Kalyanaraman B
; Hardy M
; Podsiadly R
; Cheng G
; Zielonka J
Arch Biochem Biophys
2017[Mar]; 617
(?): 38-47
PMID27590268
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In this review, some of the recent developments in probes and assay techniques
specific for superoxide (O(2)(-)) and hydrogen peroxide (H(2)O(2)) are discussed.
Over the last decade, significant progress has been made in O(2)(-) and H(2)O(2)
detection due to syntheses of new redox probes, better understanding of their
chemistry, and development of specific and sensitive assays. For superoxide
detection, hydroethidine (HE) is the most suitable probe, as the product,
2-hydroxyethidium, is specific for O(2)(-). In addition, HE-derived dimeric
products are specific for one-electron oxidants. As red-fluorescent ethidium is
always formed from HE intracellularly, chromatographic techniques are required
for detecting 2-hydroxyethidium. HE analogs, Mito-SOX and hydropropidine, exhibit
the same reaction chemistry with O(2)(-) and one-electron oxidants. Thus,
mitochondrial superoxide can be unequivocally detected using HPLC-based methods
and not by fluorescence microscopy. Aromatic boronate-based probes react
quantitatively with H(2)O(2), forming a phenolic product. However, peroxynitrite
and hypochlorite react more rapidly with boronates, forming the same product.
Using ROS-specific probes and HPLC assays, it is possible to screen chemical
libraries to discover specific inhibitors of NADPH oxidases. We hope that
rigorous detection of O(2)(-) and H(2)O(2) in different cellular compartments
will improve our understanding of their role in redox signaling.
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