Proteomic Identification of Endothelial Proteins Isolated in Situ from Atherosclerotic Aorta via Systemic Perfusion

Citations Over TimeTop 16% of 2007 papers

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.

Related Papers

• → Biotinylated erythrocytes: in vivo survival and in vitro recovery(1987)111 cited
• → Position-specific incorporation of biotinylated non-natural amino acids into a protein in a cell-free translation system(2007)23 cited
• → Biotinylation of Proteins in Solution and on Cell Surfaces(1996)10 cited
• → Biotinylated erythrocytes: in vivo survival and in vitro recovery(1987)6 cited
• → In Vivo Biotinylated scFv Fragments(2010)1 cited