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oai:repisalud.isciii.es:20.500.12105/178932024-02-10T20:01:57Zcom_20.500.12105_15322com_20.500.12105_2051col_20.500.12105_16927
Repisalud author Estévez, Mario author Padilla, Patricia author Carvalho, Leila author Martín, Lourdes author Carrapiso, Ana author Delgado, Josué 2024-02-10T20:01:57Z 2024-02-10T20:01:57Z 2019-07-20 10.1016/j.redox.2019.101277 2213-2317 Redox biology http://hdl.handle.net/10668/14322 31352127 http://hdl.handle.net/20.500.12105/17893 Carbonylation is one of the most remarkable expressions of the oxidative damage to proteins and the DNPH method the most common procedure to assess protein oxidation in biological samples. The present study was elicited by two hypotheses: i) is malondialdehyde, as a reactive dicarbonyl, able to induce the formation of allysine through a Maillard-type reaction? and ii) to which extent does the attachment of MDA to proteins interfere in the assessment of protein carbonyls using the DNPH method? Human serum albumin (HSA), human hemoglobin (HEM) and β-lactoglobulin (LAC) (5 mg/mL) were incubated with MDA (0.25 mM) for 24 h at 37 °C (HSA and HEM) or 80 °C (LAC). Results showed that MDA was unable to induce oxidative deamination of lysine residues and instead, formed stable and fluorescent adducts with proteins. Such adducts were tagged by the DNPH method, accounting for most of the protein hydrazones quantified. This interfering effect was observed in a wide range of MDA concentrations (0.05-1 mM). Being aware of its limitations, protein scientists should accurately interpret results from the DNPH method, and apply, when required, other methodologies such as chromatographic methods to detect specific primary oxidation products such as allysine. eng Allysine Carbonylation DNPH method Malondialdehyde Protein oxidation Malondialdehyde interferes with the formation and detection of primary carbonyls in oxidized proteins. research article