Use of modified CUPRAC and dinitrophenylhydrazine colorimetric methods for simultaneous measurement of oxidative protein damage and antioxidant defense against oxidation

Bayarsaikhan, Govigerel; Avan, Aslı Neslihan; Çekiç, Sema Demirci; Apak, Reşat

doi:10.1016/j.talanta.2019.06.049

Cited by 14 publications

(10 citation statements)

References 62 publications

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“…Besides the in-solution procedure, in-gel techniques based on western blotting and ELISA-type immunodetection by DNPH-tagged protein antibodies, can also be applied (Meyer et al 2012 ; Augustyniak et al 2015 ). Despite of recent optimization of the DNPH method for the assessment of protein carbonylation in food systems (Soglia et al 2016 ) and other biological samples (Colombo et al 2016 ; Bayarsaikhan et al 2019 ), this procedure suffers from lack of specificity and has been recurrently criticized for providing an unreliable estimation of the real extent of the protein oxidative damage.…”

Section: Concise Update On Methodological Approachesmentioning

confidence: 99%

Protein carbonylation in food and nutrition: a concise update

2021

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Protein oxidation is a topic of indisputable scientific interest given the impact of oxidized proteins on food quality and safety. Carbonylation is regarded as one of the most notable post-translational modifications in proteins and yet, this reaction and its consequences are poorly understood. From a mechanistic perspective, primary protein carbonyls (i.e. α-aminoadipic and γ-glutamic semialdehydes) have been linked to radical-mediated oxidative stress, but recent studies emphasize the role alternative carbonylation pathways linked to the Maillard reaction. Secondary protein carbonyls are introduced in proteins via covalent linkage of lipid carbonyls (i.e. protein-bound malondialdehyde). The high reactivity of protein carbonyls in foods and other biological systems indicates the intricate chemistry of these species and urges further research to provide insight into these molecular mechanisms and pathways. In particular, protein carbonyls are involved in the formation of aberrant and dysfunctional protein aggregates, undergo further oxidation to yield carboxylic acids of biological relevance and establish interactions with other biomolecules such as oxidizing lipids and phytochemicals. From a methodological perspective, the routine dinitrophenylhydrazine (DNPH) method is criticized not only for the lack of accuracy and consistency but also authors typically perform a poor interpretation of DNPH results, which leads to misleading conclusions. From a practical perspective, the biological relevance of protein carbonyls in the field of food science and nutrition is still a topic of debate. Though the implication of carbonylation on impaired protein functionality and poor protein digestibility is generally recognized, the underlying mechanism of such connections requires further clarification. From a medical perspective, protein carbonyls are highlighted as markers of protein oxidation, oxidative stress and disease. Yet, the specific role of specific protein carbonyls in the onset of particular biological impairments needs further investigations. Recent studies indicates that regardless of the origin (in vivo or dietary) protein carbonyls may act as signalling molecules which activate not only the endogenous antioxidant defences but also implicate the immune system. The present paper concisely reviews the most recent advances in this topic to identify, when applicable, potential fields of interest for future studies.

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Section: Concise Update On Methodological Approachesmentioning

confidence: 99%

Protein carbonylation in food and nutrition: a concise update

2021

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“…The CUPRAC assay was performed as previously described [24], with minor modifications. Briefly, 100 µL sample was withdrawn from the incubated mixtures, and protein content was precipitated using 40 µL 0.6M trichloroacetic acid solution to determine AGEs formed on BSA.…”

Section: Cuprac Measurement Of Ages In Protein Samplesmentioning

confidence: 99%

“…The CUPRAC assay was successfully applied to measure antioxidant activity of foods and bioactive compounds, while also showing its potency for determining oxidative damage of DNA and proteins using various techniques such as electrochemical or optical methods [21][22][23][24]. In the latter case, CUPRAC-reactive products generated by protein oxidation (in short time interval) were not detected in the precipitated form of albumin, but were present in the supernatant.…”

Section: Optimization Of Cuprac Assays For Detecting Agesmentioning

confidence: 99%

“…Moreover, it can be applied not only for measuring AGE biomarkers but possibly also for drug discovery regarding diseases associated with AGEs. For example, the CU-PRAC method has also been shown to simultaneously measure oxidative protein damage and inhibit antioxidant compound activity [24]. Therefore, it can be applied further to test the effects of bioactive compounds against toxic AGEs.…”

Section: Ascorbic Acid (Aa)-induced Alterations In Agesmentioning

confidence: 99%

“…The CUPRAC method is a globally used optical method and its principle is based on measuring the absorbance of the Cu (I)-Nc complex generated by the reduction reaction of Cu (II)-Nc, using a wavelength of 450 nm [21]. This method was developed by a research group led by Professor Resat Apak and has been successfully applied to measure the antioxidant activity of bioactive compounds in bio-logical samples or foods, while showing its potency for determining oxidative damage in DNA and proteins [22][23][24]. Furthermore, this system has remarkable merits; for example, the assay works considerably well at physiological pH (approximately 7.0), it uses stable reagents, and because of its low redox potential, CUPRAC reacts with a wide range of compounds, including thiols, mono-and polyphenols, carbonyls, and imidazole ringcontaining compounds [25,26].…”

Section: Introductionmentioning

confidence: 99%

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CUPRAC-Reactive Advanced Glycation End Products as Prognostic Markers of Human Acute Myocardial Infarction

Bayarsaikhan

et al. 2021

Antioxidants

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Cardiovascular disorders, especially acute coronary syndromes, are among the leading causes of mortality worldwide, and advanced glycation end products (AGEs) are associated with cardiovascular disease and serve as biomarkers for diagnosis and prediction. In this study, we investigated the utility of AGEs as prognostic biomarkers for acute myocardial infarction (AMI). We measured AGEs in serum samples of AMI patients (N = 27) using the cupric ion reducing antioxidant capacity (CUPRAC) method on days 0, 2, 14, 30, and 90 after AMI, and the correlation of serum AGE concentration and post-AMI duration was determined using Spearman’s correlation analysis. Compared to total serum protein, the level of CUPRAC reactive AGEs was increased from 0.9 to 2.1 times between 0–90 days after AMI incident. Furthermore, the glycation pattern and Spearman’s correlation analysis revealed four dominant patterns of AGE concentration changes in AMI patients: stable AGE levels (straight line with no peak), continuous increase, single peak pattern, and multimodal pattern (two or more peaks). In conclusion, CUPRAC-reactive AGEs can be developed as a potential prognostic biomarker for AMI through long-term clinical studies.

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