Involvement of the oxygen storage protein myoglobin in muscle damage under oxidative stress.
Myoglobin (Mb), the muscular oxygen reservoir, was shown to possess peroxidative reactivity in presence of H2O2 leading to oxidation of isolated cellular proteins like myosin. The objective of this study was to investigate the peroxidative effect of Mb/H2O2 on proteins in intact myofibrils (MF). Incubation of chicken leg MF in isotonic, pH 7.3 buffer at 37 degrees C in the presence of Mb (30 microM) and H2O2 (200 microM), resulted in aggregation of MF material as inspected under light microscope. SDS-PAGE analysis revealed presence of high molecular weight aggregates at the expense of myosin heavy chains, but not actin. This crosslinking was unaffected by S-S reducing agents. Continuous low flow (0.03-3.00 microM/minute), produced by glucose oxidase and glucose, was more active than bolus H2O2 addition in myosin crosslinking in MF material. Hemin which may be released from Mb under oxidative stress, was more active than Mb as a trigger of MF peroxidative aggregation. Calcium-ATPase activity of crosslinked MF was considerably lost. These findings suggest that Mb/H2O2 may lead to oxidation of neighbouring muscular protein thereby jeopardize their functioning thus explaining muscular malfunction under oxidative stress.
Kamin-Belsky
N
N
Sackler Faculty of Medicine, Tel-Aviv University, Israel.
Tomashov
R
R
Arav
R
R
Shaklai
N
N
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D000252:Calcium-Transporting ATPases / Q000378:metabolism; D002645:Chickens; D006427:Hemin / Q000378:metabolism / Q000494:pharmacology; D006861:Hydrogen Peroxide / Q000633:toxicity*; D018482:Muscle, Skeletal / Q000293:injuries / Q000502:physiology* / Q000648:ultrastructure; D009210:Myofibrils / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D009211:Myoglobin / Q000378:metabolism*; D018384:Oxidative Stress
D009211:Myoglobin; D006427:Hemin; D006861:Hydrogen Peroxide; D000251:Adenosine Triphosphatases; D000252:Calcium-Transporting ATPases