Membrane oxidation assay--a novel lipoxygenase-based evaluation of membrane oxidizability.
A membrane oxidation assay is presented which uses isolated erythrocyte membranes ("ghosts") alipoxygenase as a selective catalyst for the transfer of oxygen to cis-cis-1,4-pentadiene-moieties. The latter are, for instance, present in linoleic and arachidonic acids, both of which are integral parts of membranes. These non-conjugated double bonds represent energetically favorable sites for oxidative attack and therefore, may be rearranged and partially consumed during oxidative stress. Consequently, the measurement of oxygen consumption in the course of the lipoxygenase-mediated oxidation provides a tool for the quick and reliable determination of such double bonds. Significant inter-individual differences have been noted in 11 subjects, which also correlate to the total radical antioxidant parameter (TRAP) values obtained. This assay will be helpful in the assessment of oxidizable structures in erythrocyte membranes that may be diminished as a consequence of oxidative damage suffered by an individual. In conclusion, a simple and rapid assay for the assessment of the oxidizability of erythrocyte membranes is presented complementing the TRAP assay for plasma antioxidative status.
Lukacs
Zoltan
Z
Department of Pediatrics, University of Hamburg, Martinistr. 52, 20246 Hamburg, Germany. lukacs@uke.uni-hamburg.de
Gieseking
Jutta
J
Finckh
Barbara
B
Kohlschütter
Alfried
A
D000975:Antioxidants / Q000378:metabolism; D016718:Arachidonic Acid / Q000737:chemistry; D001774:Blood Chemical Analysis / Q000379:methods*; D002462:Cell Membrane / Q000378:metabolism*; D002784:Cholesterol / Q000378:metabolism; D004910:Erythrocyte Membrane / Q000378:metabolism; D005609:Free Radicals; D006801:Humans; D008084:Lipoxygenase / Q000378:metabolism*; D010100:Oxygen / Q000378:metabolism*; D010101:Oxygen Consumption; D013997:Time Factors
D000975:Antioxidants; D005609:Free Radicals; D016718:Arachidonic Acid; D002784:Cholesterol; D008084:Lipoxygenase; D010100:Oxygen