Effects of oxyradicals on oxymyoglobin. Deoxygenation, haem removal and iron release.
We have examined the effects of O2-derived free radicals on oxymyoglobin, the myocardial intracellular protein involved in the storage and transport of O2. The oxyradicals generated by the xanthine/xanthine oxidase system decreased the concentration of oxymyoglobin. Based on the decreases in absorbance peaks at 581 nm and 415 nm it is estimated that out of a 10 nmol decrease in oxymyoglobin, 5 nmol appears to be oxidized to ferrimyoglobin (deoxygenation), while haem was removed from the other 5 nmol of haem protein. These processes were inhibited by both catalase alone and superoxide dismutase in combination with catalase, but not by either superoxide dismutase alone or deferoxamine. These results suggest that among H2O2, OH. and O2.-, only H2O2 causes the removal of haem and the oxidation of oxymyoglobin. Furthermore, the oxyradicals also released 3 microM free iron from oxymyoglobin, which is at least 5-fold less than the 15 nmol loss of oxymyoglobin. The loss of oxymyoglobin also preceded the release of free iron. These results indicate that oxymyoglobin oxidation and haem removal occur before the removal of free iron. Thus myoglobin appears to be highly susceptible to free radical attack, and this may represent yet another mechanism of free radical-mediated cellular injury.
Prasad
M R
MR
Department of Surgery, University of Connecticut School of Medicine, Farmington 06302.
Engelman
R M
RM
Jones
R M
RM
Das
D K
DK
D000818:Animals; D005290:Ferric Compounds; D005609:Free Radicals; D006418:Heme / Q000378:metabolism; D006736:Horses; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D006878:Hydroxides / Q000494:pharmacology*; D066298:In Vitro Techniques; D007501:Iron / Q000378:metabolism; D009211:Myoglobin; D010084:Oxidation-Reduction; D013057:Spectrum Analysis; D013481:Superoxides / Q000494:pharmacology*; D013997:Time Factors
D005290:Ferric Compounds; D005609:Free Radicals; D006878:Hydroxides; D009211:Myoglobin; C017595:oxymyoglobin; D013481:Superoxides; D006418:Heme; D006861:Hydrogen Peroxide; D007501:Iron