Carbonyl proteins

Reinheckel T et al. Adaptation of protein carbonyl detection to the requirements of proteome analysis demonstrated for hypoxia/reoxygenation in isolated rat liver mitochondria. Arch Biochem Biophys 2000 376 59-65. 

Ahn, B., Rhee, S.G., and Stadtman, E.R. (1987) Use of fluorescein hydrazide and fluorescein thiosemicar-bazide reagents for the fluorometric determination of protein carbonyl groups and for the detection of oxidized proteins on polyacrylamide gels. Anal. Biocbem. 161, 245-257. 

Chaudhuri, A.R., de Waal, E.M., Pierce, A., Van Remmen, H., Ward, W.F., and Richardson, A. (2007) Detection of protein carbonyls in aging liver tissue A fluorescence-based proteomic approach. Mech. Ageing Dev. 127(11), 849-861. 

Winterbourn, C.C., and Buss, I.H. (1999) Protein carbonyl measurement by enzyme-linked immunosorbent assay. Meth. Enzymol. 300, 106-111. 

Protein peroxidation Modified tyrosines GC/MS, HPLC, immunoassays Protein carbonyls Atomic absorption spectroscopy, fluorescence spectroscopy, HPLC... 

In earlier studies the in vitro transition metal-catalyzed oxidation of proteins and the interaction of proteins with free radicals have been studied. In 1983, Levine [1] showed that the oxidative inactivation of enzymes and the oxidative modification of proteins resulted in the formation of protein carbonyl derivatives. These derivatives easily react with dinitrophenyl-hydrazine (DNPH) to form protein hydrazones, which were used for the detection of protein carbonyl content. Using this method and spin-trapping with PBN, it has been demonstrated [2,3] that protein oxidation and inactivation of glutamine synthetase (a key enzyme in the regulation of amino acid metabolism and the brain L-glutamate and y-aminobutyric acid levels) were sharply enhanced during ischemia- and reperfusion-induced injury in gerbil brain. 

There are contradictory data on the effects of dietary ascorbic acid on free radical-mediated damage in animals. Barja et al. [65] found that the administration of 660mg/kg vitamin C to guinea pigs for 5 weeks significantly decreased the levels of protein carbonyls and lipid peroxidation products. On the other hand, the administration of 500mg/kg vitamin C to... 

Overproduction of free radicals by erythrocytes and leukocytes and iron overload result in a sharp increase in free radical damage in T1 patients. Thus, Livrea et al. [385] found a twofold increase in the levels of conjugated dienes, MDA, and protein carbonyls with respect to control in serum from 42 (3-thalassemic patients. Simultaneously, there was a decrease in the content of antioxidant vitamins C (44%) and E (42%). It was suggested that the iron-induced liver damage in thalassemia may play a major role in the depletion of antioxidant vitamins. Plasma thiobarbituric acid-reactive substances (TBARS) and conjugated dienes were elevated in (3-thalassemic children compared to controls together with compensatory increase in SOD activity [386]. The development of lipid peroxidation in thalassemic erythrocytes probably depends on a decrease in reduced glutathione level and decreased catalase activity [387]. 

Figure 18.9 Production of protein carbonyls. (From Dalle-Donne et al., 2003. Copyright 2003, with permission from Elsevier.)...

Dalle-Donne, I., Giustarini, D., Colombo, R., Rossi, R. and Milzani, A. (2003) Protein carbonylation in human diseases, Trends Mol. Med., 9, 164-176. 

In these spectra, the protein has been regenerated with retinal specifically 13 C labeled at positions 11,12 and 13, and in each case the retinal resonance exhibits a sharp centerband at the isotropic chemical shift and is flanked by rotational sidebands. Other lines in the spectrum are the natural-abundance 13C resonances of the protein carbonyls ca 175 ppm) and aliphatic carbons (0-100 ppm). Contributions from the Ammonyx-LO detergent in these spectra are seen in the different intensities in the 0-100 ppm region. Ammonyx-LO does not exhibit NMR resonances above 100 ppm. Spectra of the 9-cis pigment isorhodopsin are similar. Table 38 summarizes the isotropic chemical shifts from the solid-state NMR spectra of rhodopsin regenerated with retinal13 C labeled at each position along... 

SWNTs (10 or 40pg/mouse) Male C57BL/6 mice Single intrapharyngeal instillation 7 days, 28 days, mtDNA damage was accompanied by changes in and 60 days aortic mitochondrial glutathione and protein carbonyl levels Li et al. (2007)... 

There are a number of findings suggesting that agents that facilitate elimination of protein carbonyls (by either proteolytic elimination or by enzymically mediated chemical reduction) may suppress neurodegenerative conditions in model systems (Botella et al., 2004). Consequently, as carnosine may also react with protein carbonyls, it is theoretically possible that it could suppress formation and /the reactivity of protein carbonyls in the brain. Whether carnosine participates in carbonyl reductase activity has not been investigated but it is also a reasonable speculation. 

Hipkiss, A. R. and Brownson, C. (2000). Carnosine reacts with protein carbonyl groups Another possible role for the anti-ageing peptide Biogerontology 1, 217-223. 

Hipkiss, A. R., Brownson, C., and Carrier, M. J. (2001). Carnosine, the anti-ageing, antioxidant dipeptide, may react with protein carbonyl groups. Mech. Ageing Dev. 122, 1431-1445. 

The aim of the present study was to evaluate the effect of LLLT on oxidative markers in serum and tissue biopsies of healing ulcers before and after the 8th session of an LLLT course of chronic leg ulcer treatment. Oxidative damage was assessed in terms of lipid peroxidation reflected by serum malondialdehyde (MDA) level, protein oxidation was measured in terms of tissue protein carbonyls (PCb), and DNA damage was measured in terms of DNA fragmentation. Antioxidative activity was estimated by measuring activity of SOD, GPX and CAT enzymes. 

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