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Journal of Lipid Research, Vol 32, 349-358, Copyright © 1991 by Lipid Research, Inc.
L Cominacini, U Garbin, A Davoli, R Micciolo, O Bosello, G Gaviraghi, LA Scuro and AM Pastorino
When human low density lipoprotein (LDL) obtained from 10 volunteers was
incubated in air at 37 degrees C in the presence of various concentrations
of copper, an increase in fluorescence was observed with emission maximum
at 430 nm when excitation was performed at 360 nm. The fluorescence
increase was inhibited by ethylenediamine tetraacetic acid and by
4-methyl-2,6-di-tert-butylphenol. The fluorescence was found to be tightly
bound to the protein moiety. Furthermore, Cu2+ modification of LDL was
associated with a decrease in the reactive amino groups of apolipoprotein B
and in the uptake of the lipoprotein by rabbit fibroblasts. Under our
conditions, the fluorescence increase showed two consecutive periods; an
inhibition period during which the fluorescence increased only weakly, and
a propagation period with a rapid increase in fluorescence that was linear
for at least 5 h. Both periods were influenced by copper concentration. The
study also shows that the extent of fluorescence generated upon LDL
oxidation varied greatly in the volunteers. Thus, while the results
demonstrate that the fluorescence increase may likely monitor the extent of
the apoB derivatization, the calculation of the fluorescence development
rate of the propagation period together with the duration of the inhibition
period may constitute a quantitative measurement of the susceptibility of
apoB to be derivatized.
ARTICLES
A simple test for predisposition to LDL oxidation based on the fluorescence development during copper-catalyzed oxidative modification
Department of Internal Medicine, University of Verona, Italy.
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