Protein carbonylation detection methods: A comparison

Alomari, Esra'a; Bruno, Stefano; Ronda, Luca; Paredi, Gianluca; Bettati, Stefano; Mozzarelli, Andrea

doi:10.1016/j.dib.2018.06.088

Cited by 24 publications

(19 citation statements)

References 11 publications

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“…Based on the established ferroptosis mechanism as illustrated in Figure 4A , sorafenib caused the inhibition of system xCT (cystine/glutamate antiporter) that typically mediates the exchange of extracellular L-cystine and intracellular L-glutamate across the cellular plasma membrane, leading to GSH depletion, GPX-4 inactivation, and the promotion of LPO resulting in ferroptosis. 55 Disulfide bond, a chemical bond that responds to glutathione reduction, can consume GSH and promote ferroptosis 56. SPION can increase intracellular iron concentration from low pH (pH 4.6) of late lysosome/endosomes, which can promote the Fenton reaction to ferroptosis 57.…”

Section: Resultsmentioning

confidence: 99%

Mitochondrial membrane anchored photosensitive nano-device for lipid hydroperoxides burst and inducing ferroptosis to surmount therapy-resistant cancer

et al. 2019

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Rationale: Ferroptosis is a regulated process of cell death caused by iron-dependent accumulation of lipid hydroperoxides (LPO). It is sensitive to epithelial-to-mesenchymal transition (EMT) cells, a well-known therapy-resistant state of cancer. Previous studies on nanomaterials did not investigate the immense value of ferroptosis therapy (FT) in epithelial cell carcinoma during EMT. Herein, we describe an EMT-specific nanodevice for a comprehensive FT strategy involving LPO burst.Methods: Mitochondrial membrane anchored oxidation/reduction response and Fenton-Reaction-Accelerable magnetic nanophotosensitizer complex self-assemblies loading sorafenib (CSO-SS-Cy7-Hex/SPION/Srfn) were constructed in this study for LPO produced to overcome the therapy-resistant state of cancer. Both in vitro and in vivo experiments were performed using breast cancer cells to investigate the anti-tumor efficacy of the complex self-assemblies.Results: The nano-device enriched the tumor sites by magnetic targeting of enhanced permeability and retention effects (EPR), which were disassembled by the redox response under high levels of ROS and GSH in FT cells. Superparamagnetic iron oxide nanoparticles (SPION) released Fe2+ and Fe3+ in the acidic environment of lysosomes, and the NIR photosensitizer Cy7-Hex anchored to the mitochondrial membrane, combined sorafenib (Srfn) leading to LPO burst, which was accumulated ~18-fold of treatment group in breast cancer cells. In vivo pharmacodynamic test results showed that this nanodevice with small particle size and high cytotoxicity increased Srfn circulation and shortened the period of epithelial cancer treatment.Conclusion: Ferroptosis therapy had a successful effect on EMT cells. These findings have great potential in the treatment of therapy-resistant epithelial cell carcinomas.

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Section: Resultsmentioning

confidence: 99%

Mitochondrial membrane anchored photosensitive nano-device for lipid hydroperoxides burst and inducing ferroptosis to surmount therapy-resistant cancer

et al. 2019

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“…Requena et al [6] regarded secondary carbonylation as a negligible event limited to rat liver samples in which carbonyls may also be introduced by glycation and lipid peroxidation products. In their recent review, Alomari et al [29] refer to the ‘Levine’ DNPH method as a valid biomarker of oxidatively stressed proteins. Relevant papers in the field are discriminated between those in which the complex nature of protein carbonyls is emphasized and the validity of the DNPH method as a precise technique for assessing oxidized proteins is questioned [25,[30], [31], [32]] and those in which such complexity is overlooked and protein hydrazones are assumed to mainly reflect primary oxidation of amino acid residues in proteins [33,34].…”

Section: Resultsmentioning

confidence: 99%

Malondialdehyde interferes with the formation and detection of primary carbonyls in oxidized proteins

et al. 2019

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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.

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“…8), but this assumes that carbonyls are the only reducible species, which may not always be correct (187). The methods available for detecting carbonyls have recently been reviewed (188).…”

Section: Detection and Quantification Of Productsmentioning

confidence: 99%

Detection, identification, and quantification of oxidative protein modifications

Hawkins

Davies

2019

Journal of Biological Chemistry

313

209

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Edited by Ruma Banerjee Exposure of biological molecules to oxidants is inevitable and therefore commonplace. Oxidative stress in cells arises from both external agents and endogenous processes that generate reactive species, either purposely (e.g. during pathogen killing or enzymatic reactions) or accidentally (e.g. exposure to radiation, pollutants, drugs, or chemicals). As proteins are highly abundant and react rapidly with many oxidants, they are highly susceptible to, and major targets of, oxidative damage. This can result in changes to protein structure, function, and turnover and to loss or (occasional) gain of activity. Accumulation of oxidatively-modified proteins, due to either increased generation or decreased removal, has been associated with both aging and multiple diseases. Different oxidants generate a broad, and sometimes characteristic, spectrum of post-translational modifications. The kinetics (rates) of damage formation also vary dramatically. There is a pressing need for reliable and robust methods that can detect, identify, and quantify the products formed on amino acids, peptides, and proteins, especially in complex systems. This review summarizes several advances in our understanding of this complex chemistry and highlights methods that are available to detect oxidative modifications-at the amino acid, peptide, or protein level-and their nature, quantity, and position within a peptide sequence. Although considerable progress has been made in the development and application of new techniques, it is clear that further development is required to fully assess the relative importance of protein oxidation and to determine whether an oxidation is a cause, or merely a consequence, of injurious processes.

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Protein carbonylation detection methods: A comparison

Cited by 24 publications

References 11 publications

Mitochondrial membrane anchored photosensitive nano-device for lipid hydroperoxides burst and inducing ferroptosis to surmount therapy-resistant cancer

Mitochondrial membrane anchored photosensitive nano-device for lipid hydroperoxides burst and inducing ferroptosis to surmount therapy-resistant cancer

Malondialdehyde interferes with the formation and detection of primary carbonyls in oxidized proteins

Detection, identification, and quantification of oxidative protein modifications

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