title: Proteomic identification of endothelial proteins isolated in situ from atherosclerotic aorta via systemic perfusion.
abstract: The functional and structural alterations of vascular endothelium contribute to the initiation, progression, and complications of atherosclerotic plaque formation, but limited information is known about the molecular composition and pathways underlying pathological changes during atherosclerosis. We have developed an affinity proteomic strategy for in situ isolation and differential mapping of vascular endothelial proteins in normal and atherosclerotic aorta tissues. The selective labeling was carried out by perfusion of the blood vessels with an active biotin reagent for covalent modification of accessible vascular endothelial proteins. The biotinylated proteins were then enriched by streptavidin affinity chromatography, separated by SDS-PAGE, and subsequently characterized by LC-MS/MS. The described procedure led to the identification of 454 distinct proteins in normal and atherosclerotic aorta tissues. A majority of the proteins are plasma membrane associated and extracellular matrix proteins, and 81 showed altered expressions in atherosclerotic aorta tissue. The differentially expressed proteins are involved in immune and inflammatory responses, cell adhesion, and lipid metabolism. The method provides a new avenue for investigating the endothelial dysfunction and development of atherosclerosis.
authors.affiliation: Biological Technologies, Cardiovascular and Metabolic Diseases, Wyeth Research, Cambridge, Massachusetts 02140, USA.
authors.forename: Jiang
authors.initials: J
authors.lastname: Wu
authors.forename: Wei
authors.initials: W
authors.lastname: Liu
authors.forename: Eric
authors.initials: E
authors.lastname: Sousa
authors.forename: Yongchang
authors.initials: Y
authors.lastname: Qiu
authors.forename: Debra D
authors.initials: DD
authors.lastname: Pittman
authors.forename: Vasu
authors.initials: V
authors.lastname: Maganti
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Feldman
authors.forename: Davinder
authors.initials: D
authors.lastname: Gill
authors.forename: Zhijian
authors.initials: Z
authors.lastname: Lu
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Dorner
authors.forename: Robert
authors.initials: R
authors.lastname: Schaub
authors.forename: Xiang-Yang
authors.initials: XY
authors.lastname: Tan
mesh_terms: D000818:Animals; D001013:Aorta, Thoracic / Q000737:chemistry* / Q000378:metabolism / Q000473:pathology; D050197:Atherosclerosis / Q000378:metabolism* / Q000473:pathology; D001710:Biotin; D004591:Electrophoresis, Polyacrylamide Gel; D004730:Endothelium, Vascular / Q000737:chemistry* / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D010477:Perfusion; D011506:Proteins / Q000032:analysis* / Q000145:classification / Q000378:metabolism; D040901:Proteomics; D013194:Staining and Labeling
chemical_list: D011506:Proteins; D001710:Biotin