Aleksandra Grzelakowska; Balaraman Kalyanaraman; Jacek Zielonka

DOI:

Abstract

Peroxynitrite (ONOO‒/ONOOH) is a short-lived but highly reactive species that is formed in the diffusion-controlled reaction between nitric oxide and the superoxide radical anion. It can oxidize certain biomolecules and has been considered as a key cellular oxidant formed under various pathophysiological conditions. It is crucial to selectively detect and quantify ONOO– to determine its role in biological processes. In this review, we discuss various approaches used to detect ONOO‒ in cell-free and cellular systems with the major emphasis on small-molecule chemical probes. We review the chemical principles and mechanisms responsible for the formation of the detectable products, and plausible limitations of the probes. We recommend the use of boronate-based chemical probes for ONOO‒, as they react directly and rapidly with ONOO–, they produce minor but ONOO‒‒specific products, and the reaction kinetics and mechanism have been rigorously characterized. Specific experimental approaches and protocols for the detection of ONOO– in cell-free, cellular, and in vivo systems using boronate-based molecular probes are provided (as shown in BOX 1, BOX 2, BOX 3, BOX 4, BOX 5, BOX 6).

Keywords

peroxynitrite ; molecular ; ; bioluminescence ; chromatography ; biomarkers

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