Comparison of protein oxidation and aldehyde formation during oxidative stress in isolated mitochondria.
Oxidative stress is known to cause oxidative protein modification and the generation of reactive aldehydes derived from lipid peroxidation. Extent and kinetics of both processes were investigated during oxidative damage of isolated rat liver mitochondria treated with iron/ascorbate. The monofunctional aldehydes 4-hydroxynonenal (4-HNE), n-hexanal, n-pentanal, n-nonanal, n-heptanal, 2-octenal, 4-hydroxydecenal as well as thiobarbituric acid reactive substances (TBARS) were detected. The kinetics of aldehyde generation showed a lag-phase preceding an exponential increase. In contrast, oxidative protein modification, assessed as 2,4-dinitrophenylhydrazine (DNPH) reactive protein-bound carbonyls, continuously increased without detectable lag-phase. Western blot analysis confirmed these findings but did not allow the identification of individual proteins preferentially oxidized. Protein modification by 4-HNE, determined by immunoblotting, was in parallel to the formation of this aldehyde determined by HPLC. These results suggest that protein oxidation occurs during the time of functional decline of mitochondria, i.e. in the lag-phase of lipid peroxidation. This protein modification seems not to be caused by 4-HNE.
Reinheckel
T
T
Department of Surgery, Otto-von-Guericke, University Magdeburg, Germany. Reinheckel@medizin.uni-magdeburg.de
Noack
H
H
Lorenz
S
S
Wiswedel
I
I
Augustin
W
W
D000447:Aldehydes / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D000818:Animals; D000906:Antibodies; D015153:Blotting, Western; D002845:Chromatography; D005227:Fatty Acids / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism; D007700:Kinetics; D015227:Lipid Peroxidation; D008930:Mitochondria, Liver / Q000378:metabolism*; D018384:Oxidative Stress; D010659:Phenylhydrazines / Q000276:immunology / Q000378:metabolism; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D013053:Spectrophotometry; D017392:Thiobarbituric Acid Reactive Substances / Q000378:metabolism
D000447:Aldehydes; D000906:Antibodies; D005227:Fatty Acids; D010659:Phenylhydrazines; D011506:Proteins; D017392:Thiobarbituric Acid Reactive Substances; C004787:2,4-dinitrophenylhydrazine; C010463:n-hexanal; D005978:Glutathione; C027576:4-hydroxy-2-nonenal