Lipid oxidation in biological membranes: Electron transfer proteins as initiators of lipid autoxidation

doi:10.1016/0003-9861(75)90036-3

Archives of Biochemistry and Biophysics

Volume 171, Issue 1, November 1975, Pages 292-304

https://doi.org/10.1016/0003-9861(75)90036-3 Get rights and content

Abstract

Biological lipid autoxidation has been studied in a model system composed of sonicated phospholipids as substrate and electron transfer proteins found in membranes as possible catalysts. Heme compounds, flavoproteins, and iron-sulfur proteins were examined for their ability to initiate lipid autoxidation. Among many heme compounds tested, the most active were hematin ⩾microperoxidase ⪢ methemoglobin > cytochrome c. With fresh preparations of phospholipids, reaction rates (nanomoles of oxygen/minute nanomoles of heme) ranged from 5 (cytochrome c) to 350 (hematin). Only the oxidized heme compounds were active as catalysts. Reduced heme compounds, flavoproteins and riboflavin were inactive. In the presence of heme compounds, aged preparations of sonicated phospholipids were much more rapidly oxidized than fresh preparations. They also had a higher content of fatty acid hydroperoxides as judged from their characteristic diene absorption peak at 234 nm. This observation agrees with the postulated mechanism of lipid autoxidation by heme compounds, namely, homolytic scission of preformed fatty acid hydroperoxides. Iron-sulfur proteins were also active as initiators of lipid autoxidation when destabilized in the presence of an appropriate iron chelator (o-phenanthroline or 2,2′-bipyridine) or a chaotropic ion. Oxygen uptake rates (nanomoles of oxygen/minute × milligrams of protein) varied from about 200 for an iron-sulfur protein isolated from complex I to about 5500 for Clostridium pasteurianum ferredoxin. However, per nanomole of labile sulfide, the rates for all active iron-sulfur proteins were 4–7 nmol of oxygen/min × nmol of labile sulfide.

Superoxide-generating systems did not initiate lipid autoxidation, nor did erythrocuprein inhibit the autoxidations induced by heme compounds or ferredoxin. However, lipid oxidations induced by two other iron-sulfur proteins were partially inhibited by erythrocuprein. It is concluded that in the above system Superoxide anion is neither an initiator nor an obligatory intermediate of lipid autoxidation.

References (42)

Y. Hatefi et al.
Arch. Biochem. Biophys
(1970)
W.G. Hanstein et al.
Arch. Biochem. Biophys
(1970)
F. Haurowitz et al.
J. Biol. Chem
(1941)
A.L. Tappel
Arch. Biochem. Biophys
(1953)
A.L. Tappel
J. Biol. Chem
(1955)
A.L. Tappel et al.
Arch. Biochem. Biophys
(1959)
F.P. Simon et al.
J. Biol. Chem
(1944)
S.E. Lewis et al.
Biochim. Biophys. Acta
(1963)
Y. Nakamura et al.
J. Lipid Res
(1971)
J.A. Fee et al.
Biochem. Biophys. Res. Commun
(1972)

T.C. Pederson et al.

Biochem. Biophys. Res. Commun

(1972)

T.C. Pederson et al.

Biochem. Biophys. Res. Commun

(1973)

Y. Hatefi et al.

Biochem. Biophys. Res. Commun

(1967)

W.R. Fisher et al.

J. Biol. Chem

(1973)

S. Fleischer et al.

T. Omura et al.

J. Biol. Chem

(1964)

R.L. Heath et al.

Arch. Biochem. Biophys

(1968)

J.M. McCord et al.

J. Biol. Chem

(1969)

A.L. Tappel et al.

J. Biol. Chem

(1952)

I.D. Desai et al.

J. Lipid Res

(1963)

Y. Hatefi et al.

J. Biol. Chem

(1969)

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^☆: Supported by USPHS Grant No. CA-13609. This article is Paper III of a series; Paper II appeared in Arch. Biochem. Biophys., 138, 87–95 (1970).

²: Present address: Institute for Molecular Biology, Austrian Academy of Sciences, Wasagasse 9, A1090 Vienna, Austria.

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Lipid oxidation in biological membranes: Electron transfer proteins as initiators of lipid autoxidation☆

Abstract

Arch. Biochem. Biophys

Arch. Biochem. Biophys

J. Biol. Chem

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