Abstract
Liposomes comprised of liver microsomal phospholipids and radioactive phosphatidylcholine or phosphatidylethanolamine as tracers were incubated with isolated liver microsomal NADPH-cytochrome P-450 reductase, NADPH and ADP-EDTA-chelated iron ions, a system which stimulates peroxidation of unsaturated fatty acids of phospholipids. Phospholipids and their reaction products were extracted and chromatographed on HPLC.
Phosphatidylcholine and phosphatidylethanolamine considerably decreased after 30 min incubation, depending on the enzyme and NADPH as measured by UV absorbance and radioactivity. However, neither a lysophospholipid peak nor a lysophospholipid-like peak were detectable. We suggest that lysophospholipid formation during microsomal lipid peroxidation is exclusively due to phospholipase A2 and not due to peroxidative breakdown of the unsaturated fatty acid in the β-position of glycerol.
References
Au AM, Chan PH, Fishman RA (1985) Stimulation of phospholipase A2 activity by oxygen-derived free radicals in isolated brain capillaries. J Cell Biochem 27: 449–453
Benedetti A, Casini AF, Ferrali M, Comporti M (1979) Extraction and partial characterization of dialysable products originating from the peroxidation of liver microsomal lipids and inhibiting microsomal glucose 6-phosphatase activity. Biochem Pharmacol 28: 2909–2918
Crawford CG, Plattner RD, Sessa DJ, Rackis JJ (1980) Separation of oxidized and unoxidized molecular species of phosphatidylcholine by high pressure liquid chromatography. Lipids 15: 91–94
Esterbauer H (1982) Aldehydic products of lipid peroxidation. In: McBrien DCH, Slater TF (eds) Free radicals, lipid peroxidation and cancer. Academic Press Inc, London, pp 101–128
Hughes H, Smith CV, Horning EC, Mitchell JR (1983) High-performance liquid chromatography and gas chromatography — mass spectrometry determination of specific lipid peroxidation products in vivo. Anal Biochem 130: 431–436
Kappus H (1985) Lipid peroxidation: mechanisms, analysis, enzymology and biological relevance. In: Sies H (ed) Oxidative stress. Academic Press Inc, London, pp 273–310
Kappus H, Kieczka H, Scheulen M, Remmer H (1977) Molecular aspects of catechol and pyrogallol inhibition of liver microsomal lipid peroxidation stimulated by ferrous ion-ADP-complexes or by carbon tetrachloride. Naunyn-Schmiedeberg's Arch Pharmacol 300: 179–187
Morehouse LA, Thomas CE, Aust SD (1984) Superoxide generation by NADPH-cytochrome P-450 reductase: The effect of iron chelators and the role of superoxide in microsomal lipid peroxidation. Arch Biochem Biophys 232: 366–377
Niehaus jr WG (1978) A proposed role of superoxide anion as a biological nucleophile in the deesterification of phospholipids. Bioorg Chem 7: 77–84
Omura T, Takesue S (1970) A new method for simultaneous purification of cytochrome b5 and NADPH-cytochrome c reductase from rat liver microsomes. J Biochem 67: 249–257
Pederson TC, Buege JA, Aust SD (1973) Microsomal electron transport. J Biol Chem 248: 7134–7141
Poli G, Dianzani MU, Cheeseman KH, Slater TF, Lang J, Esterbauer H (1985) Separation and characterization of the aldehydic products of lipid peroxidation stimulated by carbon tetrachloride or ADP-iron in isolated rat hepatocytes and rat liver microsomal suspensions. Biochem J 227: 629–638
Porter NA, Wolf RA, Weenen H (1980) The free radical oxidation of polyunsaturated lecithins. Lipids 15: 163–167
Remmer H, Greim H, Schenkman JB, Estabrook RW (1967) Methods for the elevation of hepatic microsomal mixed function oxidase levels and cytochrome P-450. Meth Enzymol 10: 703–708
Roders MK, Glende jr EA, Recknagel RO (1977) Prelytic damage of red cells in filtrates from peroxidizing microsomes. Science 196: 1221–1222
Scheulen ME, Kappus H, Thyssen D, Schmidt CG (1981) Redox cycling of Fe(III)-bleomycin by NADPH-cytochrome P-450 reductase. Biochem Pharmacol 30: 3385–3388
Schulze RM, Kappus H (1980) Lysis of erythrocytes as a result of microsomal lipid peroxidation induced by CCl4 or FeCl2. Res Commun Chem Pathol Pharmacol 27: 129–137
Sevanian A, Stein RA, Mead JF (1981) Metabolism of epoxidized phosphatidylcholine by phospholipase A2 and epoxide hydrolase. Lipids 16: 781–789
Tanaka Y, Mashino K, Inoue K, Nojima S (1983) Mechanism of human erythrocyte hemolysis induced by short-chain phosphatidylcholines and lysophosphatidylcholine. J Biochem 94: 833–840
Terao J, Hirota Y, Kawakatsu M, Matsushita S (1981) Structural analysis of hydroperoxides formed by oxidation of phosphatidylcholine with singlet oxygen. Lipids 427–432
Terao J, Asano I, Matsushita S (1984) High-performance liquid chromatographic determination of phospholipid peroxidation products of rat liver after carbon tetrachloride administration. Arch Biochem Biophys 235: 326–333
Ungemach FR (1985) Plasma membrane damage of hepatocytes following lipid peroxidation: involvement of phospholipase A2. In: Poli G, Cheeseman KH, Dianzani MU, Slater TF (eds) Free radicals in liver injury. IRL Press, Oxford, pp 127–134
Weglicki WB, Dickens BF, Mak IT (1984) Enhanced lysosomal phospholipid degradation and lysophospholipid production due to free radicals. Biochem Biophys Res Commun 124: 229–235
Weltzien HU (1979) Cytolytic and membrane-perturbing properties of lysophosphatidylcholine. Biochim Biophys Acta 559: 259–287
Yamamoto Y, Niki E, Kamiya A, Shimasaki H (1984) Oxidation of lipids. 7. Oxidation of phosphatidylcholines in homogenous solution and in water dispersion. Biochim Biophys Acta 795: 332–340
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Dedicated to Professor Dr. med. Herbert Remmer on the occasion of his 65th birthday
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Kostrucha, J., Kappus, H. No evidence for lysophospholipid formation during peroxidation of phospholipids by NADPH-cytochrome P-450 reductase and iron ions. Arch Toxicol 60, 170–173 (1987). https://doi.org/10.1007/BF00296974
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DOI: https://doi.org/10.1007/BF00296974