Mapping oxidations of apolipoprotein B-100 in human low-density lipoprotein by liquid chromatography-tandem mass spectrometry.
Human low-density lipoprotein (LDL) is a major cholesterol carrier in blood. Elevated concentration of low-density lipoprotein, especially when oxidized, is a risk factor for atherosclerosis and other cardiac inflammatory diseases. Past research has connected free radical initiated oxidations of LDL with the formation of atherosclerotic lesions and plaque in the arterial wall. The role of LDL protein in the associated diseases is still poorly understood, partially due to a lack of structural information. In this study, LDL was oxidized by hydroxyl radical. The oxidized protein was then delipidated and subjected to trypsin digestion. Peptides derived from trypsin digestion were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Identification of modified peptide sequences was achieved by a database search against apo B-100 protein sequences using the SEQUEST algorithm. At different hydroxyl radical concentrations, oxidation products of tyrosine, tryptophan, phenylalanine, proline, and lysine were identified. Oxidized amino acid residues are likely located on the exterior of the LDL particle in contact with the aqueous environment or directly bound to the free radical permeable lipid layer. These modifications provided insight for understanding the native conformation of apo B-100 in LDL particles. The presence of some natural variants at the protein level was also confirmed in our study.
Chakraborty
Sourav
S
Department of Chemistry, University of New Orleans, New Orleans, LA 70148, USA.
Cai
Yang
Y
Tarr
Matthew A
MA
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry; D053299:Apolipoprotein B-100 / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D017665:Hydroxyl Radical / Q000737:chemistry; D008077:Lipoproteins, LDL / Q000737:chemistry*; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D010449:Peptide Mapping; D010455:Peptides / Q000737:chemistry; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014357:Trypsin / Q000378:metabolism
D000596:Amino Acids; D053299:Apolipoprotein B-100; D008077:Lipoproteins, LDL; D010455:Peptides; D017665:Hydroxyl Radical; D014357:Trypsin