Metmyoglobin promotes arachidonic acid peroxidation at acid pH.
The ability of metmyoglobin and other heme proteins to promote peroxidation of arachidonic acid under acidic conditions was investigated. Incubation of metmyoglobin with arachidonic acid resulted in a pH-dependent increase in lipid peroxidation as measured by the formation of thiobarbituric acid reactive products and oxygen consumption. Increased peroxidation was observed at pH levels below 6.0, reaching a plateau between pH 5.5 and 5.0. At comparable heme concentrations, metmyoglobin was more efficient than oxymyoglobin, methemoglobin, or ferricytochrome c in promoting arachidonic acid peroxidation. Metmyoglobin also promoted peroxidation of 1-palmityl-2-arachidonyl phosphatidylcholine and methylarachidonate but at significantly lower rates than arachidonic acid. Addition of fatty acid-free albumin inhibited arachidonic acid peroxidation in a molar ratio of 6 to 1 (arachidonic acid:albumin). Both ionic and non-ionic detergents inhibited metmyoglobin-dependent arachidonic acid peroxidation under acidic conditions. The anti-oxidants butylated hydroxytoluene and nordihydroguaiaretic acid and low molecular weight compounds with reduced sulfhydryl groups inhibited the reaction. However, mannitol, benzoic acid, and deferoxamine were without significant effect. Visible absorption spectra of metmyoglobin following reaction with arachidonic acid showed minimal changes consistent with a low level of degradation of the heme protein during the reaction. These observations support the hypothesis that metmyoglobin and other heme proteins can promote significant peroxidation of unsaturated fatty acids under conditions of mildly acidic pH such as may occur at sites of inflammation and during myocardial ischemia and reperfusion. This may be the result of enhanced aggregation of the fatty acid and/or interaction of the fatty acid with heme under acidic conditions.
Fantone
J
J
Department of Pathology, University of Michigan Medical School, Ann Arbor 48109-0602.
Jester
S
S
Loomis
T
T
D000975:Antioxidants; D016718:Arachidonic Acid; D001095:Arachidonic Acids / Q000378:metabolism*; D005609:Free Radicals; D006420:Hemeproteins / Q000494:pharmacology*; D006863:Hydrogen-Ion Concentration; D015227:Lipid Peroxidation / Q000187:drug effects; D008315:Malondialdehyde / Q000096:biosynthesis; D008786:Metmyoglobin / Q000494:pharmacology*; D010101:Oxygen Consumption; D012710:Serum Albumin, Bovine / Q000378:metabolism
D000975:Antioxidants; D001095:Arachidonic Acids; D005609:Free Radicals; D006420:Hemeproteins; D008786:Metmyoglobin; D012710:Serum Albumin, Bovine; D016718:Arachidonic Acid; D008315:Malondialdehyde