pH-dependent regulation of myeloperoxidase activity.
The balance between peroxidase and chlorinating activities of myeloperoxidase (MPO) is very important for the enhancement of antimicrobial action and prevention of damage caused by hypochlorite. In the present paper, the peroxidase and chlorinating activities have been studied at various pH values. The possibility of using neutrophil protein solution for the evaluation of MPO activity has been demonstrated. It is shown that at neutral pH MPO had higher affinity to peroxidase substrate guaiacol: at pH 7.4, chloride ions did not compete with guaiacol up to the concentration of 150 mM. At acidic pH, chlorinating activity of MPO dominates: only hypochlorite production can be detected at equal chloride and guaiacol concentrations of 15 mM. However, horseradish peroxidase does not exhibit any difference in activity in the presence of chloride ions even at acidic pH values. It was demonstrated by MALDI-TOF mass-spectrometry that the amount of hypochlorite produced is sufficient to modify phospholipids (with formation of Cl- and Br-hydrins and lyso-derivatives) only at acidic pH (5.0). Thus, in the presence of phenolic peroxidase substrate, MPO chlorinating activity can be displayed at acidic pH only. It can lead to elimination of hypochlorite production in normal tissues at neutral pH (7.4) and its enhancement in phagosomes where the pH range is 4.7-6.0.
Vlasova
I I
II
Research Institute of Physico-Chemical Medicine, Moscow, 119992, Russia. Irina-Vlasova@newmail.ru
Arnhold
J
J
Osipov
A N
AN
Panasenko
O M
OM
D002713:Chlorine / Q000378:metabolism; D004789:Enzyme Activation; D006219:Halogens / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D006997:Hypochlorous Acid / Q000378:metabolism; D008081:Liposomes / Q000378:metabolism; D009504:Neutrophils / Q000201:enzymology / Q000378:metabolism; D017672:Nitrogen Compounds / Q000494:pharmacology; D009195:Peroxidase / Q000378:metabolism*; D010743:Phospholipids / Q000378:metabolism
D006219:Halogens; D008081:Liposomes; D017672:Nitrogen Compounds; D010743:Phospholipids; D002713:Chlorine; D006997:Hypochlorous Acid; D006861:Hydrogen Peroxide; D009195:Peroxidase