title: Identification and characterization of oxylipid-protein and peptide conjugates by mass spectrometry.
abstract: The modification of proteins by reactive products of lipid peroxidation is associated with a large number of diseases and biological aging; thus, methods that enable the characterization of oxylipid-protein and/or peptide conjugates are highly in demand. This unit outlines a chemical labeling approach to identifying and characterizing proteins modified by lipid peroxidation products. It also outlines two approaches for mass spectrometry-based identification and detailed characterization of oxylipid conjugates. The first combines chemical labeling of oxylipid-protein conjugates using an aldehyde-specific biotinylation reagent, electrophoretic separation, and mass spectrometry-based identification of the biotinylated proteins. In the second approach, protein extracts are treated with the aldehyde-specific reagent, proteolyzed using trypsin, and the biotinylated peptides are enriched using immobilized monomeric avidin. The enriched peptide fractions are submitted to tandem mass spectrometry for determining the peptide sequence information, site of the modification, and chemical nature of the oxylipid.
authors.affiliation: Oregon State University, Department of Chemistry, Corvallis, Oregon, USA.
authors.forename: Woon-Gye
authors.initials: WG
authors.lastname: Chung
authors.forename: Claudia S
authors.initials: CS
authors.lastname: Maier
mesh_terms: D004591:Electrophoresis, Polyacrylamide Gel; D015227:Lipid Peroxidation; D058635:Lipid-Linked Proteins / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification; D010446:Peptide Fragments / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification; D010449:Peptide Mapping / Q000379:methods*; D010455:Peptides / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053719:Tandem Mass Spectrometry
chemical_list: D058635:Lipid-Linked Proteins; D010446:Peptide Fragments; D010455:Peptides