title: Human apo-lipoprotein B from normal plasma contains oxidised peptides.
abstract: Oxidation of low density lipoprotein (LDL) may be atherogenic, but radical-initiated oxidation of its apoprotein B-100 (apoB) has been little studied. Transition metal ions iron and copper are candidates for mediating radical oxidation of LDL in vivo. Therefore, we studied the copper-ion-induced oxidation of apoB in human LDL. Using HPLC methods developed in our recent work, we studied the destruction of native and the generation of six oxidised amino acids; we also assessed the release of peptides from the LDL particle by FPLC. We observed time-dependent losses of apoB histidine, lysine and glycine. Long-lived reactive species, the reductant DOPA, and the oxidant hydroperoxides of valine and leucine (measured as hydroxides after reduction), were generated. Their relative abundance (mol/mol of parent amino acid) was DOPA > o- and m-tyrosine > dityrosine, valine-hydroxides, leucine hydroxides. Low molecular weight fragments were also released from the LDL in a time-dependent manner, contained hydroperoxides sensitive to GSH peroxidase, and generated radicals on reaction with iron-EDTA. The fragments contained peptides active in the quinone redox cycling procedure, comprising 0.25% of the supplied LDL amino acids. Characteristic peptides were present in each FPLC fraction containing the fragments, as judged by further HPLC fractionation. Some fragments were present in the unoxidised LDL preparations, and when these were largely removed by FPLC, copper oxidation could still generate fragments, suggesting that those present in the starting material might indicate prior oxidation. Concordantly, we found that fresh plasma LDL apoB contained 43% of total plasma protein-bound oxidised amino acids, and with the same relative abundance. We conclude that plasma proteins including apoB are subject to physiological oxidation, similar to that inflicted by copper ions; the latter may contribute to intimal LDL oxidation, which could be the source of oxidised plasma apoB.
authors.affiliation: Cell Biology Group, Heart Research Institute, Sydney, NSW, Australia.
authors.forename: D
authors.initials: D
authors.lastname: Bruce
authors.forename: S
authors.initials: S
authors.lastname: Fu
authors.forename: S
authors.initials: S
authors.lastname: Armstrong
authors.forename: R T
authors.initials: RT
authors.lastname: Dean
mesh_terms: D000596:Amino Acids / Q000032:analysis; D001055:Apolipoproteins B / Q000097:blood* / Q000737:chemistry* / Q000528:radiation effects; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004295:Dihydroxyphenylalanine; D004307:Dose-Response Relationship, Radiation; D005978:Glutathione; D006801:Humans; D017665:Hydroxyl Radical; D010084:Oxidation-Reduction; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D000596:Amino Acids; D001055:Apolipoproteins B; D010446:Peptide Fragments; D017665:Hydroxyl Radical; D004295:Dihydroxyphenylalanine; D005978:Glutathione