Determination of plasma free 3‐nitrotyrosine and tyrosine by reversed‐phase liquid chromatography with 4‐fluoro‐7‐nitrobenzofurazan derivatization

Zhang, Wei Zheng; Lang, Charles H.; Kaye, David M.

doi:10.1002/bmc.750

Cited by 25 publications

(21 citation statements)

References 20 publications

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“…As this developed and validated the method for food samples, it indicates that 3NT can be used as a protein nitration marker. Due to the lack of literature data dealing with the formation of 3NT in food samples, recently published data dealing with the formation of 3NT in biological samples were searched to determine the consistency of our results (Tsikas & Duncan, ; Weber et al, ; Zhang, Lang, & Kaye, ).…”

Section: Resultsmentioning

confidence: 99%

Investigation of the effect of sodium nitrite on protein oxidation markers in food protein suspensions

Özyurt

Ötleş

2020

J Food Biochem

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The aim of this study is to investigate the effect of sodium nitrite (NaNO2) on protein oxidation and the use of 3‐nitrotyrosine (3NT) as a protein oxidation marker in suspensions of the food protein. Food proteins, namely bovine serum albumin, casein, and myofibrillar protein, were suspended in 100 mM sodium phosphate buffer and nitrated with 25 µM iron (III) chloride, 2.5 mM hydrogen peroxide, and 150 mg/kg NaNO2 at 37°C for a period of 24 hr. The food protein suspensions were analyzed at different sampling periods for the loss of tryptophan (TRY) residues as well as the formation of Schiff bases (SBs), protein carbonyls, 3NT, and dityrosine (DT). It was found that NaNO2 has pro‐oxidant activity in NaNO2‐added food protein suspensions due to the increased amounts of SBs, protein carbonyls, 3NT, and DT as well as decreased TRY fluorescence. Positive correlations between the 3NT and other protein oxidation markers except for TRY fluorescence were found in NaNO2‐added food protein suspensions. In conclusion, these findings on the detection of 3NT indicate that it might be a useful tool as a new protein oxidation biomarker in food samples. Practical applications Sodium nitrite (NaNO2) supports protein oxidation in different food protein suspension. 3‐nitrotyrosine (3NT) was found in food protein suspensions and can be a potential biomarker for protein nitration in food sample due to potential relationship between 3NT and other oxidation markers. The results showed that this study has formed novel insight into interaction between NaNO2 and food proteins and indicate that it might affect the food quality and its nutritional value. Moreover, the relationship between protein oxidation/nitration and food quality as well as the comprehension of the scientific and technological meaning of these phenomena has been hindered because of the lack of knowledge about the basic chemistry behind the protein oxidation and nitration pathways. For the clarification of these phenomena, further studies are still needed.

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Section: Resultsmentioning

confidence: 99%

Investigation of the effect of sodium nitrite on protein oxidation markers in food protein suspensions

Özyurt

Ötleş

2020

J Food Biochem

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“…Derivatization with 4-fluoro-7-nitrobenzeno-2-oxa-1,3-diazole was performed. A 10 µl aliquot was used for HPLC quantification as described [28]. …”

Section: Methodsmentioning

confidence: 99%

Neonatal Asphyxia Induces the Nitration of Cardiac Myosin Light Chain 2 That is Associated with Cardiac Systolic Dysfunction

Doroszko

Polewicz

Cadete

et al. 2010

Shock

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Background Hypoxia followed by reoxygenation (H-R) observed during perinatal asphyxia is a serious complication with a high mortality and morbidity rate which may cause adverse cardiovascular effects in neonates. Our aim was to determine if oxidative stress related to H-R induces peroxynitrite-dependent modifications of the cardiac contractile protein, myosin regulatory light chain 2 (MLC2) and whether this is associated with development of cardiac systolic dysfunction. Methods and Results Twelve newborn piglets were acutely instrumented for hemodynamic monitoring and randomized to a control group ventilated with only atmospheric air or to the H-R study group, exposed to alveolar normocapnic hypoxia followed by reoxygenation. Afterwards, animals were euthanized and the hearts harvested for biochemical analyses. Systolic function as well as cardiac MLC2 levels decreased in H-R animals, whereas nitrates and nitrotyrosine levels increased. Negative correlations between nitrates, nitrotyrosine and MLC2 levels were observed. Moreover, H-R induced nitration of two tyrosine residues within the MLC2 protein. Similarly, in vitro exposure of MLC2 to peroxynitrite resulted in the nitration of tyrosine, which increased the susceptibility of MLC2 to subsequent degradation by matrix metalloproteinase-2 (MMP-2). Substitution of this tyrosine with phenylalanine prevented the MMP-2 dependent degradation of MLC2. In addition, a large decrease in MLC2 phosphorylation due to H-R was observed. Conclusions 1. Oxidative stress related to asphyxia induces nitration of cardiac MLC2 protein and thus increases its degradation. This and a large decrease in MLC2 phosphorylation contribute to the development of systolic dysfunction. 2. Inhibition of MLC2 nitration and/or direct inhibition of its degradation by MMP-2 could be potential therapeutic targets aiming at reduction of myocardial damage during resuscitation of asphyxiated newborns.

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“…Precolumn derivatization of NT with NBD-F for determination of it in plasma, has been reported by some studies [13,21,22]. In the present study, precolumn derivatization of NT with NBD-F has been applied for the measurements of plasma free NT in a relatively large number of patients as compared to the former studies.…”

Section: Figure 1 Bivariate Correlations Between Plasma Myeloperoxidmentioning

confidence: 77%

Association between Plasma Myeloperoxidase and Free 3-Nitrotyrosine Levels in Patients with Coronary Artery Disease

Pourfarzam¹,

Ahmad²,

Sarrafzadegan³

et al. 2013

IJCM

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Objective: Myeloperoxidase (MPO) is an inflammatory enzyme that is mainly released by activated neutrophils and monocytes. 3-nitrotyrosine (NT) is a stable inflammatory end product of MPO that is produced through nitrosylation of free and protein-bound tyrosines. Determination of the exact levels of free NT is technically a challenging matter. Also, there is limited information about the relationship between MPO and free NT levels and elevation of them in the plasma of patients with coronary artery disease (CAD). Therefore, we sought to determine the exact level of plasma free NT with a simple and exquisite technique in CAD patients. Methods: This study included 50 stable angina, 50 unstable angina patients, and 50 control subjects. Plasma MPO concentration was measured with an immunoassay method. Plasma free NT level was determined by a modified HPLC-fluorescence method. Lipid profile, high sensitivity C-reactive protein (hsCRP) and other clinical risk factors of patients were also assigned. Results: Plasma level of free NT was efficiently measured by the HPLC-fluorescence method. Plasma levels of MPO and NT were significantly higher in patients with stable and unstable CAD than in control subjects (P < 0.001). There was a significant correlation between the two substances in CAD patients (P < 0.001). Conclusions: We determined plasma free NT levels with a sensitive HPLC-fluorescence method with some modifications in a clinical scale. Plasma levels of MPO and NT were profoundly elevated in CAD patients. The significant relationships of the two substances and elevation of them may have useful clinical implication in patients with stable and unstable CAD.

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Determination of plasma free 3‐nitrotyrosine and tyrosine by reversed‐phase liquid chromatography with 4‐fluoro‐7‐nitrobenzofurazan derivatization

Cited by 25 publications

References 20 publications

Investigation of the effect of sodium nitrite on protein oxidation markers in food protein suspensions

Investigation of the effect of sodium nitrite on protein oxidation markers in food protein suspensions

Neonatal Asphyxia Induces the Nitration of Cardiac Myosin Light Chain 2 That is Associated with Cardiac Systolic Dysfunction

Association between Plasma Myeloperoxidase and Free 3-Nitrotyrosine Levels in Patients with Coronary Artery Disease

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