2011
DOI: 10.1002/psc.1298
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Epitope motif of an anti‐nitrotyrosine antibody specific for tyrosine‐nitrated peptides revealed by a combination of affinity approaches and mass spectrometry

Abstract: Nitration of tyrosine residues has been shown to be an important oxidative modification in proteins and has been suggested to play a role in several diseases such as atherosclerosis, asthma, lung and neurodegenerative diseases. Detection of nitrated proteins has been mainly based on the use of nitrotyrosine-specific antibodies. In contrast, only a small number of nitration sites in proteins have been unequivocally identified by MS. We have used a monoclonal 3-NT-specific antibody, and have synthesized a series… Show more

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Cited by 21 publications
(35 citation statements)
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“…As pointed out in recent studies [33,46], the characterization of affinities and binding specificities of monoclonal 3-NT-specific antibodies are of crucial importance and should be ascertained using synthetic tyrosine-nitrated peptides; high antibody affinities were found for nitrotyrosine peptides containing basic (Lys, Arg) amino acid residues in the vicinity of the nitrated tyrosine ( Figure 5). Corresponding tyrosine-nitrated peptides may also find important application as biomarkers of oxidative modifications.…”
Section: Discussionmentioning
confidence: 99%
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“…As pointed out in recent studies [33,46], the characterization of affinities and binding specificities of monoclonal 3-NT-specific antibodies are of crucial importance and should be ascertained using synthetic tyrosine-nitrated peptides; high antibody affinities were found for nitrotyrosine peptides containing basic (Lys, Arg) amino acid residues in the vicinity of the nitrated tyrosine ( Figure 5). Corresponding tyrosine-nitrated peptides may also find important application as biomarkers of oxidative modifications.…”
Section: Discussionmentioning
confidence: 99%
“…Determination of antibody/ protein association kinetics was performed by extracting the data from the sensor signals using the monomolecular growth model [47]. Determinations of dissociation constants was performed by plotting the pseudo first-order kinetic constants for two independent channels verss concentration, and applying linear regression for K D =k off k on -1 for nitration sites, comprising cationic amino acids in the vicinity of nitrated tyrosine residues (Table 2) [46]. The possible role of sequence and structural environment for nitration has been discussed by several authors [11,23,33,36,40,[46][47][48].…”
Section: Site Selectivity Of Tyrosine Nitration In Proteinsmentioning
confidence: 99%
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“…3.2 ) the use of anti-3-nitro-tyrosine-specifi c antibodies has been essential for mass spectrometric identifi cation of tyrosine nitrations using proteolytic affi nity-MS. Poor information regarding the specifi cities of anti-3-nitrotyrosine antibodies has been reported before [ 4 ]. In a previous publication we described a molecular study of the recognition specifi cities and affi nities of two commercially available, monoclonal anti-nitrotyrosine antibodies by dot-blot, ELISA, and affi nity-MS, using different 3-nitrotyrosine-containing peptides [ 36 ].…”
Section: Online Bioaffi Nity-mass Spectrometry For Molecular Recognitmentioning
confidence: 99%