Nitrotyrosine: Quantitative Analysis, Mapping in Proteins, and Biological Significance

Souza, José Maria de; Bartesaghi, Silvina; Peluffo, Gonzalo; Radí, Rafael

doi:10.1002/9780813814438.ch12

Cited by 1 publication

(1 citation statement)

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“…The detection of NT was made immunochemically using anti-protein 3-nitrotyrosine antibodies, a sensitive method that was improved and extensively used afterwards [98]. In addition, bioanalytical methods have been developed for quantitation of NT levels and have been reviewed recently elsewhere [99]. …”

Section: Peroxynitrite Footprintsmentioning

confidence: 99%

Kinetic and mechanistic considerations to assess the biological fate of peroxynitrite

Carballal

Bartesaghi

Radí

2014

Biochimica et Biophysica Acta (BBA) - General Subjects

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137

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Background Peroxynitrite, the product of the reaction between superoxide radicals and nitric oxide, is an elusive oxidant of short half-life and low steady-state concentration in biological systems; it promotes nitroxidative damage. Scope of Review We will consider kinetic and mechanistic aspects that allow rationalizing the biological fate of peroxynitrite from data obtained by a combination of methods that include fast kinetic techniques, electron paramagnetic resonance and kinetic simulations. In addition, we provide a quantitative analysis of peroxynitrite production rates and conceivable state-state levels in living systems. Major Conclusions The preferential reactions of peroxynitrite in vivo include those with carbon dioxide, thiols and metalloproteins; its homolysis represents only < 1 % of its fate. To note, carbon dioxide accounts for a significant fraction of peroxynitrite consumption leading to the formation of strong one-electron oxidants, carbonate radicals and nitrogen dioxide. On the other hand, peroxynitrite is rapidly reduced by peroxiredoxins, which represent efficient thiol-based peroxynitrite detoxification systems. Glutathione, present at mM concentration in cells and frequently considered a direct scavenger of peroxynitrite, does not react sufficiently fast with it in vivo; glutathione mainly inhibits peroxynitrite-dependent processes by reactions with secondary radicals. The detection of protein 3-nitrotyrosine, a molecular footprint, can demonstrate peroxynitrite formation in vivo. Basal peroxynitrite formation rates in cells can be estimated in the order of 0.1 to 0.5 μM s−1 and its steady-state concentration ~ 1 nM. General Significance The analysis provides a handle to predict the preferential fate and steady-state levels of peroxynitrite in living systems. This is useful to understand pathophysiological aspects and pharmacological prospects connected to peroxynitrite.

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Section: Peroxynitrite Footprintsmentioning

confidence: 99%