The mechanism of peroxidase-mediated cytotoxicity. II. Role of the heme moiety.
Various peroxidases in the presence of hydrogen peroxide and a halide ion have been shown to exert a cytolytic activity against erythrocytes and other cells. However, few studies have been done to elucidate the active site on the enzymes that is responsible for the cytotoxic activity. In addressing this question we found that boiling of horseradish peroxidase only partially abolishes its cytotoxic activity, suggesting that an intact tertiary structure of the protein may not be essential for the cytotoxic activity. This conclusion was confirmed by demonstrating that microperoxidase, hemin, and hematoheme also exert cytotoxic activity in the presence of hydrogen peroxide and iodide, the kinetics of which were identical to those obtained with the peroxidases. Fluoride, bromide, and thiocyanate could not replace iodide in any of these systems. These results indicate that the active site for the cytotoxic activity of the peroxidases is located within the heme moiety, whereas the protein portions of the enzymes affect the cytotoxic activity of the enzymes only in an indirect manner. We also tested a variety of compounds for their ability to inhibit the cytolytic reaction toward erythrocytes. We found that compounds such as thiourea, thionicotinamide, and uric acid are much more potent inhibitors of the cytolytic reaction than tyrosine and histidine. These observations support the concept that oxidative reactions rather than halogenation reactions are the primary cause of the peroxidase-mediated lysis of erythrocytes.
Lin
H
H
Department of Biochemistry, Texas Tech University Health Sciences Center, Lubbock 79430.
McFaul
S J
SJ
Brady
J C
JC
Everse
J
J
D000818:Animals; D001665:Binding Sites; D004912:Erythrocytes / Q000378:metabolism*; D006418:Heme / Q000378:metabolism*; D006461:Hemolysis; D006639:Histidine / Q000378:metabolism; D006358:Hot Temperature; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007454:Iodides / Q000494:pharmacology; D007501:Iron / Q000378:metabolism; D010544:Peroxidases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011166:Porphyrins / Q000378:metabolism; D011489:Protein Denaturation; D011817:Rabbits
D007454:Iodides; D011166:Porphyrins; D006418:Heme; D006639:Histidine; D006861:Hydrogen Peroxide; D007501:Iron; D010544:Peroxidases; C021799:microperoxidase