The effect of high-density lipoproteins on the formation of lipid/protein conjugates during in vitro oxidation of low-density lipoprotein.
High-density lipoprotein (HDL) incubated with low-density lipoprotein (LDL) under oxidising conditions has previously been reported to decrease the accumulation of lipid peroxides on LDL and to diminish the biological effects of LDL, which would have been present had it been oxidatively modified in the absence of HDL. Thus far direct evidence that oxidative modification of LDL is diminished by HDL has, however, been lacking. We used electrospray ionisation mass spectrometry (ESI-MS) to detect 4-hydroxy-2-nonenal (HNE)-modified histidine residues of tryptic fragments of LDL which had been subject to Cu(2+) induced oxidation both in the presence and absence of human or avian HDL. HNE-modified angiotensin II was introduced into the incubation mixture as an internal standard and to check that HDL did not interfere in the detection of HNE-modified peptides non-specifically. Our results confirmed earlier reports that HNE modification of histidine occurs during the oxidation of LDL and for the first time revealed a marked attenuation of the process in the presence of human HDL with no effect on the detection of HNE-modified angiotensin II by ESI-MS. Avian HDL, which lacks the anti-oxidative enzyme paraoxonase, did not affect the formation of apo B adducts. Our findings therefore suggest that covalent linkage of lipid peroxidation products to LDL protein as well as the accumulation of lipid peroxides on LDL is diminished in the presence of HDL containing paraoxonase.
Sangvanich
Polkit
P
Michael Barber Centre for Mass Spectrometry, UMIST, Manchester M60 1QD, UK.
Mackness
Bharti
B
Gaskell
Simon J
SJ
Durrington
Paul
P
Mackness
Michael
M
D000447:Aldehydes / Q000032:analysis; D006639:Histidine / Q000737:chemistry; D006801:Humans; D015227:Lipid Peroxidation; D008075:Lipoproteins, HDL / Q000494:pharmacology*; D008077:Lipoproteins, LDL / Q000737:chemistry* / Q000378:metabolism*; D010084:Oxidation-Reduction; D021241:Spectrometry, Mass, Electrospray Ionization
D000447:Aldehydes; D008075:Lipoproteins, HDL; D008077:Lipoproteins, LDL; C099252:oxidized low density lipoprotein; D006639:Histidine; C027576:4-hydroxy-2-nonenal