title: A Comparison of Methods To Enhance Protein Detection of Lipoproteins by Mass Spectrometry.
abstract: We sought to develop a new method to more efficiently analyze lipid-bound proteins by mass spectrometry using a combination of a lipid removal agent (LRA) that selectively targets lipid-bound proteins and a mass spectrometry compatible detergent, anionic acid labile surfactant (AALS), that is capable of eluting proteins off the LRA. This method was compared to established methods that use the lipid removal agent alone and straight proteomic analysis of human plasma after organic solvent delipidation (OSD). Plasma from healthy individuals was separated by gel filtration chromatography and prepared for mass spectrometry analysis by each of the described methods. The addition of AALS to LRA increased the overall number of proteins detected in both the high and low density lipoprotein size range, the number of peptide counts for each protein, and the overall sequence coverage. Organic solvent delipidation detected the most proteins, though with some decrease in overall protein detection and sequence coverage due to the presence of nonlipid-bound proteins. The use of LRA allows for selection and analysis of lipid-bound proteins. The addition of a mass spectrometry compatible detergent improved detection of lipid-bound proteins from human plasma using LRA.
authors.forename: Anna
authors.initials: A
authors.lastname: Heink
authors.affiliation: §Center for Lipid and Arteriosclerosis Science, Department of Pathology and Laboratory Medicine, University of Cincinnati, 2120 East Galbraith Road, Cincinnati, Ohio 45237-0507, United States.
authors.forename: W Sean
authors.initials: WS
authors.lastname: Davidson
authors.forename: Debi K
authors.initials: DK
authors.lastname: Swertfeger
authors.forename: L Jason
authors.initials: LJ
authors.lastname: Lu
authors.forename: Amy S
authors.initials: AS
authors.lastname: Shah
mesh_terms: D002850:Chromatography, Gel; D006801:Humans; D008074:Lipoproteins / Q000032:analysis*; D008297:Male; D013501:Surface-Active Agents / Q000737:chemistry; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D008074:Lipoproteins; D013501:Surface-Active Agents
keywords: detergents; lipids; lipoproteins; mass spectrometry; surfactant