Abstract
We have determined by means of a standard spectrophotometric assay that lipid oxidation occurred at a significant rate in large, multilamellar vesicles containing egg phosphatidylcholine under normal experimental conditions. We have also observed that the fluorescence intensity of the vesicle-associated probe, 1,6-diphenyl-1,3,5-hexatriene, decreased with time in vesicles containing such oxidizing lipids. The spectrophotometric data utilized to monitor lipid oxidation were found to fit an apparent first-order kinetic model. The loss of diphenylhexatriene fluorescence intensity in oxidizing liposomes was analyzed in terms of a first-order event superimposed (and thus presumably dependent) upon the ongoing formation of oxidized lipid. These and other data were used to conclude that the oxidation-induced loss of diphenylhexatriene fluorescence intensity was due to chemical modification of the fluorophore rather than to excited-state quenching or ground-state complex formation. Finally, the loss of fluorescence intensity in oxidizable membranes was found to alter drastically the ‘microviscosity’ parameter as derived from diphenylhexatriene fluorescence anisotropy and relative intensity measurements.
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