Covalent crosslinking of the heme prosthetic group to myoglobin by H2O2: toxicological implications.
It is known that treatment of myoglobin with H2O2 leads to covalent alteration of the heme prosthetic group with concomitant formation of a protein bound heme adduct and transforms myoglobin from an oxygen storage protein to an oxidase. In the current study it was shown, with the use of 14C-labeled heme reconstituted into apomyoglobin, that up to 88% of the oxidatively altered heme can be accounted for by the protein bound product. Furthermore, a partially purified preparation of the protein bound heme adduct was introduced into human fibroblasts using the method of osmotic lysis of pinosomes and found to cause cell death (40%) within 1 h, as evidenced by trypan blue exclusion. Native myoglobin introduced into cells in the same manner or extracellular treatment by the protein bound heme adduct had no effect on cell viability. The extent of cell death could be decreased (50%) by N-acetyl-L-cysteine, indicating a potential role for reactive oxygen intermediates in this process. These results show that the covalently altered myoglobin can elicit cellular damage and suggests that similar processes may occur in vivo in pathologic conditions such as that involving cardiac ischemia and reperfusion injury, where covalently altered myoglobin may form.
Osawa
Y
Y
Laboratory of Molecular Immunology, NHLBI, NIH, Bethesda, MD 20892-1760, USA.
Williams
M S
MS
D000111:Acetylcysteine / Q000494:pharmacology; D000818:Animals; D001059:Apoproteins / Q000737:chemistry / Q000378:metabolism*; D001665:Binding Sites; D002250:Carbon Radioisotopes; D016923:Cell Death / Q000187:drug effects; D002460:Cell Line; D003432:Cross-Linking Reagents; D005347:Fibroblasts; D006418:Heme / Q000737:chemistry / Q000378:metabolism*; D006736:Horses; D006801:Humans; D006861:Hydrogen Peroxide; D009211:Myoglobin / Q000737:chemistry / Q000378:metabolism*; D010088:Oxidoreductases / Q000378:metabolism*; D011485:Protein Binding; D011865:Radioisotope Dilution Technique
D001059:Apoproteins; D002250:Carbon Radioisotopes; D003432:Cross-Linking Reagents; D009211:Myoglobin; C010857:apomyoglobin; D006418:Heme; D006861:Hydrogen Peroxide; D010088:Oxidoreductases; D000111:Acetylcysteine