Influence of HSA and IgG on LDL oxidation studied by size-exclusion chromatography and phospholipid profiling using MALDI tandem-mass spectrometry
| Autor: | Omar Belgacem, Kurt Widhalm, Gerald Stübiger, Katharina Pock, Grazyna Sobal |
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| Rok vydání: | 2011 |
| Předmět: |
Adult
Male Size-exclusion chromatography Phospholipid Mass spectrometry Tandem mass spectrometry Biochemistry Lipid peroxidation chemistry.chemical_compound In vivo TBARS Humans Molecular Biology Phospholipids Serum Albumin Chromatography Organic Chemistry Lysophosphatidylcholines Cell Biology Middle Aged Atherosclerosis Sphingomyelins Lipoproteins LDL chemistry Immunoglobulin G Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Chromatography Gel Fatty Acids Unsaturated Female lipids (amino acids peptides and proteins) Sphingomyelin Oxidation-Reduction |
| Zdroj: | Chemistry and Physics of Lipids. 164:563-572 |
| ISSN: | 0009-3084 |
| Popis: | In the present study a direct detection approach combining size-exclusion chromatography (SEC) and matrix-assisted laser desorption/ionization quadrupole ion trap time-of-flight tandem-mass spectrometry (MALDI-QIT-TOF-MS/MS) was applied to investigate the influence of HSA and IgG on LDL oxidation in vitro. SEC analysis showed an increase of protein aggregation during LDL-oxidation that could be essentially suppressed in the presence of HSA. In parallel, lipid peroxidation measured by TBARS assay over 24h was inhibited by 95-100% in the presence of HSA but only 0-34% by IgG, respectively. MALDI phospholipid profiles showed considerable decrease of signals from PCs containing sn-2 PUFAs (18:2 or 20:4) accompanied by increase of sn-2 LPCs indicating for specific breakdown of PUFA-containing PLs during LDL-oxidation. These effects were nearly 100% inhibited in the presence of HSA but not by IgG, respectively. Among known pro-atherogenic PL species present in human plasma sphingomyelin (SM16:0) was bound in significant amounts to HSA but not IgG after incubation with oxLDL. Moreover, our investigation showed that LPCs containing SAFAs (16:0 or 18:0) were specifically bound to HSA, while those containing PUFAs (18:2 and 18:3) were preferentially associated with IgG. In summary, the presented methodology provides a promising platform for studying lipid-protein interactions in vivo. |
| Databáze: | OpenAIRE |
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