Investigation of local primary structure effects on peroxynitrite-mediated tyrosine nitration using targeted mass spectrometry

Seeley, Kent W.; Stevens, Stanley M.

doi:10.1016/j.jprot.2011.11.025

Cited by 14 publications

(9 citation statements)

References 26 publications

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“…This nitrated peptide has also been identified in other studies where similar modification site selectivity has been observed [15,20]. Our group has further described the influence of local chemical environment on tyrosine nitration selectivity [35]. …”

Section: Resultssupporting

confidence: 65%

“…Although uncommon, peptide sequences with multiple nitrotyrosine residues would require adequate fragment ion coverage to perform relative quantitation of nitration at a site-specific level. In our experience, CID-based fragmentation does not show a neutral loss or possible scrambling of the NO 2 group [35,36], yet these events should perhaps be considered during data analysis, particularly when HCD/CID parameters have been modified considerably or other dissociation methods are utilized. Extensive characterization of nitropeptide fragmentation by various dissociation methods has been previously reported by other labs [38–40].…”

Section: Resultsmentioning

confidence: 99%

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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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Section: Resultssupporting

confidence: 65%

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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show abstract

“…Instead, signals of nitrated peptides have to be related to their non‐modified homologs, respectively. Even in case of different ionization efficiencies of nitrated peptides and their non‐nitrated siblings the relative response was shown to be maintained . Comparisons of signal heights and areas for nitrated and non‐nitrated peptide siblings allowed to estimate the relative nitration propensity of either site (Fig.…”

Section: Resultsmentioning

confidence: 99%

Top‐down and bottom‐up characterization of nitrated birch pollen allergen Bet v 1a with CZE hyphenated to an Orbitrap mass spectrometer

Gusenkov

Stutz

2018

Electrophoresis

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Tyrosine (Tyr) residues of the major pollen allergen of birch Betula verrucosa, Bet v 1a, were nitrated by peroxynitrite. This modification enhances the allergenicity. Modified tyrosines were identified by analyzing intact allergen variants in combination with top‐down and bottom‐up approaches. Therefore, a laboratory‐built sheath‐liquid assisted ESI interface was applied for hyphenation of CE to an Orbitrap mass spectrometer to localize individual nitration sites. The major focus was on identification of primary nitration sites. The top‐down approach unambiguously identified Tyr 5 as the most prominent modification site. Fragments from the allergen core and the C‐terminal part carried up to three potential nitration sites, respectively. Thus, a bottom‐up approach with tryptic digest was used as a complementary strategy which allowed for the unambiguous localization of nitration sites within the respective peptides. Nitration propensity for individual Tyr residues was addressed by comparison of MS signals of nitrated peptides relative to all cognates of homolog primary sequence. Combined data identified surface exposed Tyr 5 and Tyr 66 as major nitration sites followed by less accessible Tyr 158 whereas Tyr 81, 83 and 150 possess a lower nitration tendency and are apparently modified in variants with higher nitration levels.

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“…Nitration of tyrosine residues by peroxynitrite under physiological conditions has been studied in detail [132,283]. The mechanism of the reaction in the presence of excess CO 2 at pH 7.5 is presented in Figure 5.…”

Section: Reactivity With Amino Acids Peptides and Proteinsmentioning

confidence: 99%

Reactive Inorganic Oxy‐Species of C, N, S, and P

Sharma¹

2012

Oxidation of Amino Acids, Peptides, and Proteins

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Investigation of local primary structure effects on peroxynitrite-mediated tyrosine nitration using targeted mass spectrometry

Cited by 14 publications

References 26 publications

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

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

Top‐down and bottom‐up characterization of nitrated birch pollen allergen Bet v 1a with CZE hyphenated to an Orbitrap mass spectrometer

Reactive Inorganic Oxy‐Species of C, N, S, and P

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