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A simple test for predisposition to LDL oxidation based on the fluorescence development during copper-catalyzed
L Cominacini1, U Garbin, A Davoli
1Department of Internal Medicine, University of Verona, Italy.
Journal of Lipid Research
|February 1, 1991
Summary
Copper-induced oxidation of human low-density lipoprotein (LDL) generates fluorescence, offering a new method to quantify LDL susceptibility to modification. This fluorescence monitoring can assess apolipoprotein B derivatization and LDL oxidation extent in volunteers.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Lipid Metabolism
Background:
- Low-density lipoprotein (LDL) modification, particularly oxidation, is implicated in atherosclerosis.
- Quantifying LDL oxidation susceptibility is crucial for understanding cardiovascular disease risk.
Purpose of the Study:
- To investigate copper-induced LDL oxidation and its relationship with fluorescence.
- To develop a quantitative method for measuring LDL susceptibility to apolipoprotein B (apoB) derivatization.
Main Methods:
- Incubation of human LDL with varying copper concentrations at 37°C.
- Measurement of fluorescence emission at 430 nm (excitation at 360 nm).
- Assessment of reactive amino groups and fibroblast uptake of modified LDL.
Main Results:
- Copper-induced LDL oxidation resulted in increased fluorescence, tightly bound to the protein moiety.
- Oxidation decreased reactive amino groups on apoB and reduced LDL uptake by fibroblasts.
- Fluorescence increase exhibited inhibition and propagation periods, influenced by copper concentration and varying among individuals.
Conclusions:
- Fluorescence increase serves as a reliable indicator of apoB derivatization during LDL oxidation.
- The rate of fluorescence development and inhibition period duration provide a quantitative measure of apoB susceptibility to modification.