Tyrosine-Nitrated Proteins: Proteomic and Bioanalytical Aspects.
"Nitroproteomic" is under active development, as 3-nitrotyrosine in proteins constitutes a footprint left by the reactions of nitric oxide-derived oxidants that are usually associated to oxidative stress conditions. Moreover, protein tyrosine nitration can cause structural and functional changes, which may be of pathophysiological relevance for human disease conditions. Biological protein tyrosine nitration is a free radical process involving the intermediacy of tyrosyl radicals; in spite of being a nonenzymatic process, nitration is selectively directed toward a limited subset of tyrosine residues. Precise identification and quantitation of 3-nitrotyrosine in proteins has represented a "tour de force" for researchers. Recent Advances: A small number of proteins are preferential targets of nitration (usually less than 100 proteins per proteome), contrasting with the large number of proteins modified by other post-translational modifications such as phosphorylation, acetylation, and, notably, S-nitrosation. Proteomic approaches have revealed key features of tyrosine nitration both in vivo and in vitro, including selectivity, site specificity, and effects in protein structure and function.



Identification of 3-nitrotyrosine-containing proteins and mapping nitrated residues is challenging, due to low abundance of this oxidative modification in biological samples and its unfriendly behavior in mass spectrometry (MS)-based technologies, that is, MALDI, electrospray ionization, and collision-induced dissociation.



The use of (i) classical two-dimensional electrophoresis with immunochemical detection of nitrated proteins followed by protein ID by regular MS/MS in combination with (ii) immuno-enrichment of tyrosine-nitrated peptides and (iii) identification of nitrated peptides by a MIDAS™ experiment is arising as a potent methodology to unambiguously map and quantitate tyrosine-nitrated proteins in vivo. Antioxid. Redox Signal. 26, 313-328.
1 Unidad de Bioquímica y Proteómica Analíticas, Institut Pasteur de Montevideo , Montevideo, Uruguay .|2 Departamento de Bioquímica, Facultad de Medicina, Universidad de la República , Montevideo, Uruguay .|3 Facultad de Medicina, Center for Free Radical and Biomedical Research , Universidad de la República, Montevideo, Uruguay .
Carlos
C
Batthyány
3 Facultad de Medicina, Center for Free Radical and Biomedical Research , Universidad de la República, Montevideo, Uruguay .|4 Departamento de Educación Médica, Facultad de Medicina, Universidad de la República , Montevideo, Uruguay .
Silvina
S
Bartesaghi
2 Departamento de Bioquímica, Facultad de Medicina, Universidad de la República , Montevideo, Uruguay .|3 Facultad de Medicina, Center for Free Radical and Biomedical Research , Universidad de la República, Montevideo, Uruguay .
Mauricio
M
Mastrogiovanni
1 Unidad de Bioquímica y Proteómica Analíticas, Institut Pasteur de Montevideo , Montevideo, Uruguay .
Analía
A
Lima
2 Departamento de Bioquímica, Facultad de Medicina, Universidad de la República , Montevideo, Uruguay .|3 Facultad de Medicina, Center for Free Radical and Biomedical Research , Universidad de la República, Montevideo, Uruguay .
Verónica
V
Demicheli
2 Departamento de Bioquímica, Facultad de Medicina, Universidad de la República , Montevideo, Uruguay .|3 Facultad de Medicina, Center for Free Radical and Biomedical Research , Universidad de la República, Montevideo, Uruguay .
Rafael
R
Radi
D000818:Animals; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods / Q000592:standards; D009566:Nitrates / Q000737:chemistry / Q000378:metabolism*; D015538:Nitrosation; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D020543:Proteome / Q000378:metabolism; D040901:Proteomics / Q000379:methods; D012680:Sensitivity and Specificity; D014443:Tyrosine / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism*
D009566:Nitrates; D011506:Proteins; D020543:Proteome; C002744:3-nitrotyrosine; D014443:Tyrosine
free radicals; nitration; peroxynitrite; protein oxidation; proteomics