title: Human low density lipoprotein as a target of hypochlorite generated by myeloperoxidase.
abstract: The aim of this study was to further clarify which part of human low density lipoprotein (LDL) is attacked by the MPO/H2O2/Cl- -system and which reactive oxygen species is responsible for the attack. Therefore the influence of this system on the modification of the lipid and protein moiety of LDL was studied in vitro. Using the monochlorodimedone assay it was found that HOCl is produced in micromolar quantities in the absence of LDL and is rapidly consumed by LDL in a concentration dependent manner. The consumption of HOCl was reflected in the formation of HOCl-specific epitopes on apo B-100 as determined by an antibody raised against HOCl-modified LDL. The absorbency at 234 nm was applied to measure continuously the extent of modification of LDL. The general kinetic pattern of the absorbency measurement consisted of a lag phase where no LDL modification was observed, followed by a rapid increase of absorbency and a plateau phase. Finally the absorbency decreased due to LDL precipitation. Time dependent absorption spectra indicated that this kinetic pattern is mainly caused by light scattering due to particle aggregation rather than by a specific absorption at 234 nm due to conjugated diene formation. In agreement with this finding a low rate of thiobarbituric acid reactive substances (TBArS) formation was observed after a lag phase. The aggregation of LDL occurs most likely by modification of apo B-100, which was determined fluorimetrically in terms of LDL-tryptophan destruction in presence of the MPO/H2O2/Cl(-)-system. The kinetic course of tryptophan fluorescence generally consisted of a rapid decrease leveling off into a low plateau phase. Gas chromatographic determinations of linoleic acid in LDL in presence of the MPO system showed that this polyunsaturated fatty acid (PUFA) is easily attacked by HOCl. Consistent with this finding NMR spectra of HOCl modified LDL indicated a complete disappearance of bis-allylic methylene groups. Since lipid peroxidation products only partially account for this loss of PUFAs, other reactions of HOCl with unsaturated lipids--probably chlorohydrin formation--must be involved. Summarizing, although the rate of lipid peroxidation is low, both the lipid and the protein moiety of LDL are readily modified by the MPO system. It appears that the immediate consequence of apo B-100 modification is its aggregation. It is concluded that MPO, which has been detected in atherosclerotic lesions, is able to contribute to the modification of LDL into a form recognizable for uncontrolled uptake by macrophages.
authors.affiliation: Institute of Biochemistry, University of Graz, Austria.
authors.forename: A
authors.initials: A
authors.lastname: Jerlich
authors.forename: J S
authors.initials: JS
authors.lastname: Fabjan
authors.forename: S
authors.initials: S
authors.lastname: Tschabuschnig
authors.forename: A V
authors.initials: AV
authors.lastname: Smirnova
authors.forename: L
authors.initials: L
authors.lastname: Horakova
authors.forename: M
authors.initials: M
authors.lastname: Hayn
authors.forename: H
authors.initials: H
authors.lastname: Auer
authors.forename: H
authors.initials: H
authors.lastname: Guttenberger
authors.forename: H J
authors.initials: HJ
authors.lastname: Leis
authors.forename: F
authors.initials: F
authors.lastname: Tatzber
authors.forename: G
authors.initials: G
authors.lastname: Waeg
authors.forename: R J
authors.initials: RJ
authors.lastname: Schaur
mesh_terms: D000328:Adult; D053299:Apolipoprotein B-100; D001055:Apolipoproteins B / Q000097:blood / Q000737:chemistry / Q000378:metabolism; D001161:Arteriosclerosis / Q000209:etiology; D001665:Binding Sites; D005260:Female; D005609:Free Radicals / Q000378:metabolism; D006801:Humans; D006997:Hypochlorous Acid / Q000378:metabolism*; D066298:In Vitro Techniques; D007700:Kinetics; D008077:Lipoproteins, LDL / Q000097:blood / Q000737:chemistry / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D009195:Peroxidase / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet; D017392:Thiobarbituric Acid Reactive Substances / Q000378:metabolism; D014364:Tryptophan / Q000737:chemistry
chemical_list: D053299:Apolipoprotein B-100; D001055:Apolipoproteins B; D005609:Free Radicals; D008077:Lipoproteins, LDL; D017382:Reactive Oxygen Species; D017392:Thiobarbituric Acid Reactive Substances; D006997:Hypochlorous Acid; D014364:Tryptophan; D009195:Peroxidase