In vitro low-density lipoprotein oxidation by copper or *OH/O*(2)(-): new features on carbonylation and fragmentation of apolipoprotein B during the lag phase.
The aim of our study was to evaluate the carbonylation and the carbonylated fragmentation of apolipoprotein B upon low-density lipoprotein (LDL) oxidation induced in vitro by copper and *OH/O*(2)(-) free radicals generated by gamma-radiolysis. Therefore, we developed a very sensitive Western blot immunoassay using 2,4-dinitrophenylhydrazine which allows the revelation of the apolipoprotein B carbonylation and its carbonylated fragmentation. The main results of this study show that (i) apolipoprotein B carbonylation is present during the lag phase of LDL oxidation in the two oxidative processes and (ii) apolipoprotein B carbonylated fragmentation was not detected during the lag phase of copper-oxidized LDL but was detected during the propagation phase. By contrast, apolipoprotein B carbonylated fragmentation was detected in the lag phase of *OH/O*(2)(-) oxidized LDL.
Laboratoire de Biochimie Métabolique et Clinique, Faculté de Pharmacie, Paris, France. cct@kb.inserm.fr
Stéphanie
S
Zarev
Dominique
D
Bonnefont-Rousselot
Claudine
C
Cosson
Jean-Louis
JL
Beaudeux
Jacques
J
Delattre
Monique
M
Gardès-Albert
Alain
A
Legrand
Patrice
P
Thérond
D001055:Apolipoproteins B / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism* / Q000528:radiation effects; D015153:Blotting, Western; D003300:Copper / Q000494:pharmacology*; D005720:Gamma Rays; D006801:Humans; D017665:Hydroxyl Radical / Q000494:pharmacology*; D066298:In Vitro Techniques; D008077:Lipoproteins, LDL / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism* / Q000528:radiation effects; D010084:Oxidation-Reduction; D010446:Peptide Fragments / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism / Q000528:radiation effects; D013481:Superoxides / Q000494:pharmacology*
D001055:Apolipoproteins B; D008077:Lipoproteins, LDL; D010446:Peptide Fragments; D013481:Superoxides; D017665:Hydroxyl Radical; D003300:Copper