Protein tyrosine nitration

Mattson, M. P., Goodman, Y., Luo, H., Fu, W., and Furukawa, K. (1997). Activation of NF-kappaB protects hippocampal neurons against oxidative stress-induced apoptosis Evidence for induction of manganese superoxide dismutase and suppression of peroxynitrite production and protein tyrosine nitration.. Neurosci. Res. 49, 681-697. 

Radi, R. Nitric oxide, oxidants, and protein tyrosine nitration. Proc. Natl. Acad. 

Moreover, targeted disruption (knock-down) of the eNOS gene attenuates the neuronal cell death in thiamine-deficient mice (Gibson and Zhang, 2002). eNOS knock-down but not knock-down of iNOS or nNOS leads to a reduction in protein tyrosine nitration (Beauchesne et al., 2009), suggesting a major role of eNOS as the source of nitric oxide-related nitrosative stress in thiamine deficiency. 

Monteiro HP. Signal transduction by protein tyrosine nitration competition or cooperation with tyrosine phosphorylation-depen-... 

Peluffo G, Radi R. Biochemistry of protein tyrosine nitration in cardiovascular pathology. Cardiovasc Res 2007 75 291-302. 

Astrocytes from the cerebral cortex and hippocampus express functional NMDA receptors which are implicated in both astrocyte-neuron and astrocyte-astrocyte metabolic coupling (Verkhratsky and Kirchhoff, 2007). Astrocyte NMDA receptors respond to glutamate ligands with increases in intracellular Ca (Porter and McCarthy, 1995), NO production (Mollace et al 1995) and protein tyrosine nitration (Schliess et al, 2002). Furthermore, glutamate induces astrocytic swelling due to stimulation of Ca -dependent K+ uptake that is sensitive to NMDA receptor antagonists (Bender et al., 1998). 

Exposure of cultured astrocytes to millimolar concentrations of anunonia results in oxidative stress and in protein tyrosine nitration (ScUiess et al., 2002), a process that was dependent upon stimulation of the inducible isoform of NOS (NOS-2). Astrocytic protein tyrosine nitration was also described in the brain of rats following portacaval anastomosis where tyrosine nitration of GS was described (Schliess et al., 2002). Reduced GS enzyme activity resulting from tyrosine nitration offers a cogent explanation for the consistent reports of decreased capacity for ammonia removal in the brain of animals following portacaval anastomosis. 

A dramatic selective enrichment in nitrotyrosine content was observed within apoA-I recovered from serum and human atherosclerotic lesions. The analysis of serum from sequential subjects demonstrates that the nitrotyrosine contents of apoA-I were markedly higher in individuals with cardiovascular disease. An increase in nitrotyrosine was observed in the atherosclerotic lesions and plasma of apolipoprotein A-I-deficient mice (Parastatidis et al., 2007). LC MS/MS studies confirm that nitration of apoA-I occurs specifically on tyrosine-192 (Shao et al., 2005). Similarly, LC—MS/MS methods were used to identify tyrosine residues 292 and 422 at the carboxyl terminus of the 3 chain as the principal sites of fibrinogen nitration in vivo (Parastatidis et al., 2008). Stable isotope dilution LC MRM/MS methodology will likely be employed extensively in the future to assess the role of protein tyrosine nitration in the oxidative stress associated with cardiovascular disease. 

Radi, R. Protein Tyrosine Nitration Biochemical Mechanisms and Structure Basis of Functional Effects. Acc. Chem. Res. 2013, 46, 550-559. 

I. Diaz-Moreno, J. M. Garcia-Heredia, K. Gonzalez-Arzola, A. Diaz-Quintana and M. A. De la Rosa, Recent Methodological Advances in the Analysis of Protein Tyrosine Nitration, ChemPhysChem, 2013,14, 3095. 

Fig, 5 Protein tyrosine-nitration in motomeurons from ALS mice. This experiment is described in detail by Jung et al, (r. Spinal cord samples were stained with an antibody specific for nitrotyrosine. The spinal cord samples were obtained from (A) wildtype mice receiving no treatment or ALS mice (G93A) treated with (B) vehicle (C) EUK-134 or D) EUK-8, Three animals from each group were analyzed with similar results, Motomeurons (arrows) show cytoplasmic staining indicative of protein nitration in ALS mice. Reprinted from reference 60, copyright (2001), with permission from Elsevier,... 

Enzymes modified with N -carbonyldiimidazole (CDI) include horseradish peroxidase 761 /1-lactamase after nitration and reduction,[771 lysozyme, and urease.[781 Ref. [77] describes how the tyrosine side chain of a protein was nitrated, reduced with dithionite to an amino group, and then treated with CDI or A/-(2,2,2-trifluoro-ethoxycarbonyl)imidazole to give the benzoxazolinonyl alanine moiety ... 

Greenacre, S.A., and Ischiropoulos, H. (2001) Tyrosine nitration Localisation, quantification, consequences for protein function and signal transduction. Free Radic. Res. 34, 541-581. 

Protein tyrosine residues constitute key targets for peroxynitrite-mediated nitrations. Attack of various free radicals (ONOO-, N02 ) upon tyrosine generates 3-nitrotyrosine, which can be measured immunologically or by GC/MS or HPLC techniques. The detection of 3-nitrotyrosine was considered a biomarker of peroxynitrite action in vivo. Similarly, attack of HOC1 and HOBr on tyrosine generates chlorotyro-sine and bromotyrosine, respectively, both of which are measured most accurately by GC-MS. 

Similar to peroxynitrite, ONOOCOO- reacts with many biomolecules such as uric acid [110], oxyhemoglobin [133], melatonin [135], NADH, ubiquinol Q0, and glutathione [141], Reactions of ONOOCOO with substrates in mitochondrial matrix is accompanied by protein nitration [141]. The reaction of ONOOCOO- with GSH was so rapid that glutathione inhibited tyrosine nitration by peroxynitrite in the presence of C02 [142], The formation of ONOOCOO- increased the formation of 3-nitrotyrosine and decreased the formation of 3-hydroxytyrosine probably due to the enhanced selectivity of C03 - compared to hydroxyl radicals [143],... 

Recently, Gunther et al. [41] proposed that nitric oxide may directly react with enzymes without intermediate formation of peroxynitrite. It is known that the oxidation of arachido-nic acid by prostaglandin H oxidase is mediated by the formation of enzyme tyrosyl radical (see Chapter 26). Correspondingly, it has been suggested that NO is able to react with this radical to form the tyrosine iminoxyl radical and then nitrotyrosine. Therefore, the NO-dependent nitration of protein tyrosine residue may occur without the formation of peroxynitrite or other nitrogen oxides. 

Bruijn, L. I., Beal, M. F., Becher, M. W. et al. Elevated free nitrotyrosine levels, but not protein-bound nitrotyrosine or hydroxyl radicals, throughout amyotrophic lateral sclerosis (ALS)-like disease implicate tyrosine nitration as an aberrant in vivo property of one familial ALS-linked superoxide dismutase 1 mutant. Proc. Natl Acad. Sci. U.S.A. 94 7606-7611,1997. 

Beckman, J. S., Ye, Y. Z., Anderson, P., Chen, j., Accavetti, M. A., Tarpey, M. M., and White, C. R. (1994). Extensive nitration of protein tyrosines observed in human atherosclerosis detected by immunohistochemistry. Biol. Chem. Hoppe-Seyler 375, 81-88. 

Peroxynitrite has been suggested to be formed from nitric oxide and superoxide in vivo. It is a highly reactive oxidant, and causes nitration on the aromatic ring of free tyrosine and protein tyrosine residues. It was reported that peroxynitrite induced various oxidative damage in vitro, for example LDL oxidation, lipid peroxidation, and DNA strand breakage [30]. 

There are numerous data that peroxynitrite is involved in cell death and tissue injuries in many clinical conditions. An important mechanism underlying peroxynitrite toxicity is the reaction of tyrosine nitration. Tyrosine nitration inactivates certain enzymes, as was postulated for prostacyclin (PGI2) synthase (M14), cytochrome P450 2B1 (RIO), tyrosine hydroxylase (A 14), and MnSOD (Yl). Moreover, nitration blocks tyrosine phosphorylation, and thus interferes with the tyrosine kinase signaling pathways (K18). The peroxynitrite treatment of rat liver epithelial cells stimulates mitogen-activated protein kinases p38 MAPK, JNK1/2, and ERK1/2 the mechanism of this effect awaits elucidation (S9). 

Another marker for protein oxidation is nitration of tyrosine residues, and numerous previous studies support the notion that nitrosative stress also contributes to neurodegeneration in AD (Smith et al., 1997 Tohgi et al., 1999 Castegna et al, 2003 Sultana et al., 2006b). A number of mechanisms for tyrosine nitration of protein have been proposed, and the two widely believed to exist in vivo involve formation of peroxynitrite or mediation via hemeperoxidases (Brennan et al., 2002). These mechanisms involve NO or its by-products that react with ROS (Beckman et al. 

Recently, several studies suggested that protein nitration could be a cellular signaling mechanism and is often a reversible and selective process, similar to protein phosphorylation (Aulak et al., 2004 Koeck et al., 2004). In addition, modified proteins are believed to be either degraded or subject to processes that could lead to enzymatic denitration (Gow et al., 1996 Kamisaki et al., 1998 Me et al., 2003). The latter possibility is intriguing because this would allow the process of tyrosine nitration to be reversible and thus enable a more dynamic physiological role. Protein nitration is observed under normal conditions in all tissues. In AD brain levels of nitrated proteins were found to be increased compared to that of control (Smith et al.. 

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