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Proteomic identification of endothelial proteins isolated in situ from atherosclerotic aorta via systemic perfusion.
- Source :
-
Journal of proteome research [J Proteome Res] 2007 Dec; Vol. 6 (12), pp. 4728-36. Date of Electronic Publication: 2007 Nov 13. - Publication Year :
- 2007
-
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.
- Subjects :
- Animals
Aorta, Thoracic metabolism
Aorta, Thoracic pathology
Atherosclerosis pathology
Biotin
Electrophoresis, Polyacrylamide Gel
Endothelium, Vascular metabolism
Mice
Mice, Inbred C57BL
Mice, Knockout
Proteins classification
Proteins metabolism
Staining and Labeling
Aorta, Thoracic chemistry
Atherosclerosis metabolism
Endothelium, Vascular chemistry
Perfusion
Proteins analysis
Proteomics
Subjects
Details
- Language :
- English
- ISSN :
- 1535-3893
- Volume :
- 6
- Issue :
- 12
- Database :
- MEDLINE
- Journal :
- Journal of proteome research
- Publication Type :
- Academic Journal
- Accession number :
- 17997517
- Full Text :
- https://doi.org/10.1021/pr070537c