RT Journal Article
T1 Malondialdehyde interferes with the formation and detection of primary carbonyls in oxidized proteins.
A1 Estévez, Mario
A1 Padilla, Patricia
A1 Carvalho, Leila
A1 Martín, Lourdes
A1 Carrapiso, Ana
A1 Delgado, Josué
K1 Allysine
K1 Carbonylation
K1 DNPH method
K1 Malondialdehyde
K1 Protein oxidation
AB 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.
YR 2019
FD 2019-07-20
LK http://hdl.handle.net/10668/14322
UL http://hdl.handle.net/10668/14322
LA en
DS RISalud
RD Apr 11, 2025