Adriamycin-Fe3+-induced mitochondrial protein damage with lipid peroxidation.
Exposure of mitochondria to adriamycin (ADM)-Fe3+ induced formation of thiobarbituric acid reactive substances and fluorescent substances. Butylated hydroxytoluene (BHT) and the water soluble vitamin E analogue, trolox, not only strongly inhibited fluorescence formation but also mitochondrial lipid peroxidation. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated the formation of high molecular weight proteins when mitochondria were exposed to ADM-Fe3+. A mitochondrial protein with a molecular weight of approximately 30 kDa was very sensitive to ADM-Fe3+. BHT and trolox strongly inhibited mitochondrial protein cross-linking, indicating that the protein modification was due to ADM-Fe(3+)-induced lipid peroxidation. In addition, the susceptibility of ADM-Fe(3+)-exposed mitochondrial protein to proteases was unchanged. Bovine serum albumin (BSA) inhibited ADM-Fe(3+)-induced mitochondrial lipid peroxidation. Fluorescence emmited from BSA was detected during ADM-Fe(3+)-induced mitochondrial lipid peroxidation, and BHT strongly inhibited the oxidative modification of BSA. These results suggest that the oxidative modification of mitochondrial proteins and BSA is due to ADM-Fe(3+)-induced lipid peroxidation.
Miura
T
T
Department of Biochemistry, Hokkaido Institute of Pharmaceutical Sciences, Otaru, Japan.
Muraoka
S
S
Ogiso
T
T
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D002084:Butylated Hydroxytoluene / Q000494:pharmacology; D002839:Chromans / Q000494:pharmacology; D004317:Doxorubicin / Q000037:antagonists & inhibitors / Q000633:toxicity*; D004591:Electrophoresis, Polyacrylamide Gel; D006868:Hydrolysis; D066298:In Vitro Techniques; D015227:Lipid Peroxidation / Q000187:drug effects*; D008929:Mitochondria, Heart / Q000187:drug effects* / Q000378:metabolism; D009124:Muscle Proteins / Q000737:chemistry / Q000187:drug effects*; D009942:Organometallic Compounds / Q000037:antagonists & inhibitors / Q000633:toxicity*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors*; D051381:Rats; D017208:Rats, Wistar; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D013050:Spectrometry, Fluorescence; D013438:Sulfhydryl Compounds / Q000378:metabolism; D017392:Thiobarbituric Acid Reactive Substances / Q000378:metabolism
D000975:Antioxidants; D002839:Chromans; D009124:Muscle Proteins; D009942:Organometallic Compounds; D013438:Sulfhydryl Compounds; D017392:Thiobarbituric Acid Reactive Substances; C041415:doxorubicin-iron complex; D002084:Butylated Hydroxytoluene; D012710:Serum Albumin, Bovine; D004317:Doxorubicin; D011493:Protein Kinase C; C010643:6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid