The use of cis-parinaric acid to determine lipid peroxidation in human erythrocyte membranes. Comparison of normal and sickle erythrocyte membranes.
The recently developed parinaric acid assay is shown to offer possibilities for studying peroxidation processes in biological membrane systems. Taking the human erythrocyte membrane as a model, several initiating systems were investigated, as well as the effect of residual hemoglobin in ghost membrane preparations. The effectivity of a radical generating system appeared to be strongly dependent upon whether radicals are generated at the membrane level or in the water phase. Thus, cumene hydroperoxide at concentrations of 1.0-1.5 mM was found to be a very efficient initiator of peroxidation in combination with submicromolar levels of hemin-Fe3+ as membrane-bound cofactor. In combination with cumene hydroperoxide, membrane-bound hemoglobin appeared to be about 6-times more effective in promoting peroxidation than hemoglobin in the water phase. Results comparing the behaviour of normal and sickle erythrocyte ghost suspensions in the peroxidation assay suggest that the increased oxidative stress on sickle erythrocyte membranes could be due to enhanced membrane binding of sickle hemoglobin, but also partly to a characteristically higher capability of sickle hemoglobin to promote peroxidation. The order of peroxidation-promoting capabilities that could be derived from the experiments was hemin greater than sickle hemoglobin greater than normal hemoglobin.
Department of Biochemistry, State University of Utrecht, The Netherlands.
J J
JJ
Van den Berg
F A
FA
Kuypers
J H
JH
Qju
D
D
Chiu
B
B
Lubin
B
B
Roelofsen
J A
JA
Op den Kamp
D000755:Anemia, Sickle Cell / Q000097:blood*; D001555:Benzene Derivatives / Q000494:pharmacology; D004910:Erythrocyte Membrane / Q000378:metabolism*; D005231:Fatty Acids, Unsaturated / Q000032:analysis*; D006454:Hemoglobins / Q000378:metabolism; D006801:Humans; D008054:Lipid Peroxides / Q000096:biosynthesis* / Q000097:blood
D001555:Benzene Derivatives; D005231:Fatty Acids, Unsaturated; D006454:Hemoglobins; D008054:Lipid Peroxides; C007164:cumene hydroperoxide; C013641:parinaric acid