Protein carbonylation as a major hallmark of oxidative damage: Update of analytical strategies

Abstract

Protein carbonylation, one of the most harmful irreversible oxidative protein modifications, is considered as a major hallmark of oxidative stress-related disorders. Protein carbonyl measurements are often performed to assess the extent of oxidative stress in the context of cellular damage, aging and several age-related disorders. A wide variety of analytical techniques are available to detect and quantify protein-bound carbonyls generated by metal-catalyzed oxidation, lipid peroxidation or glycation/glycoxidation. Here we review current analytical approaches for protein carbonyl detection with a special focus on mass spectrometry-based techniques. The utility of several carbonyl-derivatization reagents, enrichment protocols and especially advanced mass spectrometry techniques are compared and discussed in detail. Furthermore, the mechanisms and biology of protein carbonylation are summarized based on recent high-throughput proteomics data.

Histone retention, protein carbonylation, and lipid peroxidation in spermatozoa: Possible role in recurrent pregnancy loss

“…Collectively, proteins can be oxidized in about 35 ways, out of which many modifications involve some form of carbonylation making carbonylation as the most commonly found irreversible form of protein oxidation [Dalle-Donne et al 2003]. The probable mechanism of introducing a carbonyl group to protein involves oxidative modification of specific amino acids, namely arg, lys, pro, and thr, to aldehyde or ketone and their eventual cleavage [Berlett and Stadtman 1997;Curtis et al 2012;Fedorova et al 2014]. In addition, the peptide backbone can also be oxidatively cleaved either by the α-amidation pathway or by oxidation of glutamyl, aspartyl, and proyl chain, generating α-ketonyl derivative at the N-terminal end [Stadtman and Levine 2000].…”

“…While enzymatic reversal of over-oxidative modification has not yet been detected, the potential fate of oxidized protein is proteosome proteolytic breakdown. However, excessive oxidation and cross linking of proteins would render them resistant to proteolysis leading to the accumulation of these proteins as ubiquitinated oxidatively modified proteins [Fedorova et al 2014]. …”

Gigaspora margarita with and without its endobacterium shows adaptive responses to oxidative stress

“…Several types of ROS react with methionine, forming methionine sulfoxide, which is reduced by methionine sulfoxide reductase (Luo and Levine 2009). We previously demonstrated that under normal growth conditions, the B-line contains a higher quantity of carbonylated proteins (Vannini et al 2016), which are considered a major hallmark of oxidative damage (Fedorova et al 2014) and specifically involve methionine oxidation. The gene coding for methionine sulfoxide reductase showed a significant up-regulation in our data set upon H2O2 treatment (comp18281_c1, log2 fold change:…”

Sodium Lauryl Sulfate Stimulates the Generation of Reactive Oxygen Species through Interactions with Cell Membranes

“…Further, SLS-treated cells showed an increase of intracellular calpain activity associated with the increase of intracellular Ca 2+ concentration. The increase of intracellular ROS was abolished by the addition of BAPTA-AM, a specific chelator of Ca are oxidized proteins produced by reactions between free amino residues of proteins and reactive aldehyde compounds yielded by lipid peroxidation 6 . The sum of these facts suggest that excessive oxidative stress is one of the critical causes of rough skin.…”

“…A recent study has reported that carbonylated proteins are observed in corneocytes of dry skin in the winter season 4 . That fact was hypothesized to contribute to oxidative stress in the development of skin roughness, because carbonylated proteins are end-products of lipid peroxidation initiated by ROS 6,18 .…”