Synthesis of Peroxynitrite from Nitrite and Hydrogen Peroxide

Robinson, Kristine M.; Beckman, Joseph S.

doi:10.1016/s0076-6879(05)96019-9

Cited by 103 publications

(75 citation statements)

References 13 publications

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“…Peroxynitrite was synthesized by rapidly mixing acidified hydrogen peroxide with sodium nitrite and quenching the reaction with sodium hydroxide as previously described (70). Residual hydrogen peroxide was eliminated with manganese dioxide, and the peroxynitrite concentration was determined spectrophotometrically using e 302 = 1,700 M…”

Section: Methodsmentioning

confidence: 99%

Nitration of Hsp90 induces cell death

Franco

Ye²,

Refakis

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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Oxidative stress is a widely recognized cause of cell death associated with neurodegeneration, inflammation, and aging. Tyrosine nitration in these conditions has been reported extensively, but whether tyrosine nitration is a marker or plays a role in the cell-death processes was unknown. Here, we show that nitration of a single tyrosine residue on a small proportion of 90-kDa heat-shock protein (Hsp90), is sufficient to induce motor neuron death by the P2X7 receptor-dependent activation of the Fas pathway. Nitrotyrosine at position 33 or 56 stimulates a toxic gain of function that turns Hsp90 into a toxic protein. Using an antibody that recognizes the nitrated Hsp90, we found immunoreactivity in motor neurons of patients with amyotrophic lateral sclerosis, in an animal model of amyotrophic lateral sclerosis, and after experimental spinal cord injury. Our findings reveal that cell death can be triggered by nitration of a single protein and highlight nitrated Hsp90 as a potential target for the development of effective therapies for a large number of pathologies.apoptosis | peroxynitrite | PC12 cells

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Section: Methodsmentioning

confidence: 99%

Nitration of Hsp90 induces cell death

Franco

Ye²,

Refakis

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

122

157

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“…Employing a scaled-down version of a previously-described method [30], we synthesized 30-mL volumes using a syringe pump system to combine NaNO 2 or Na 15 NO 2 and acidified H 2 O 2 in order to produce either light or heavy ONOO − (Figure 1a). Due to ONOO − instability at low pH, concentrated NaOH was added post-reaction from a separate syringe.…”

Section: Resultsmentioning

confidence: 99%

Evaluation of a Method for Nitrotyrosine Site Identification and Relative Quantitation Using a Stable Isotope-Labeled Nitrated Spike-In Standard and High Resolution Fourier Transform MS and MS/MS Analysis

Seeley

Fertig

Dufresne

et al. 2014

IJMS

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The overproduction of reactive oxygen and nitrogen species (ROS and RNS) can have deleterious effects in the cell, including structural and possible activity-altering modifications to proteins. Peroxynitrite is one such RNS that can result in a specific protein modification, nitration of tyrosine residues to form nitrotyrosine, and to date, the identification of nitrotyrosine sites in proteins continues to be a major analytical challenge. We have developed a method by which 15N-labeled nitrotyrosine groups are generated on peptide or protein standards using stable isotope-labeled peroxynitrite (O15NOO−), and the resulting standard is mixed with representative samples in which nitrotyrosine formation is to be measured by mass spectrometry (MS). Nitropeptide MS/MS spectra are filtered using high mass accuracy Fourier transform MS (FTMS) detection of the nitrotyrosine immonium ion. Given that the nitropeptide pair is co-isolated for MS/MS fragmentation, the nitrotyrosine immonium ions (at m/z = 181 or 182) can be used for relative quantitation with negligible isotopic interference at a mass resolution of greater than 50,000 (FWHM, full width at half-maximum). Furthermore, the standard potentially allows for the increased signal of nitrotyrosine-containing peptides, thus facilitating selection for MS/MS in a data-dependent mode of acquisition. We have evaluated the methodology in terms of nitrotyrosine site identification and relative quantitation using nitrated peptide and protein standards.

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“…PN was obtained in NaNO 2 and H 2 O 2 reaction in acid water solutions with their subsequent rapid alkalization [10]. To this end 100 ml 1.2 M NaNO 2 was rapidly mixed with 100 ml H 2 O 2 (10 ml 30% H 2 O 2 , 60 ml 1 M HNO 3 , 30 ml H 2 O) and 100 ml 1.5 M NaOH was added to the mixture.…”

Section: Methodsmentioning

confidence: 99%