Synthetic Gel of Carotid Artery Plaque

Abstract

Atherosclerosis is a complex disease that affects medium and large arteries, leading to the formation and progression of plaque. In this process the proteins play an essential role and as a consequence, proteomic-based strategies examining the protein content of cells or tissues could offer a useful approach for the study of plaque proteins. Due to the heterogeneous cell composition of plaque, proteome analysis of whole lesions is difficult, besides being also complicated by the presence of plasma proteins that cannot be completely eliminated. A good way to study variations in protein expression among series of gels is to construct a synthetic gel. This type of gel is obtained by averaging the positions, shapes and optical densities of spots in a given set of gels. To be included in the synthetic gel, spots must be found in at least three gels. To obtain a profile representative of the proteome of atherosclerotic plaque, canceling its high variability, we constructed a synthetic gel using an average of ten carotid plaque samples. We then compared it with an equivalent synthetic gel constructed using ten plasma samples from the same carotid surgery patients. For the comparison of two synthetic gels (plasma/plaque) we could discriminate plasma proteins from plaque proteins. Besides analysis of spots common to plasma, the synthetic gel is useful to detect spots exclusive to plaque, thus simplifying a very complex mixture.Atherosclerosis is a chronic disease that affects medium and large arteries. The process originates from the interaction between cells of the arterial wall, lipoproteins and inflammatory cells (1), leading to the development of complex lesions or plaque that protrude into the arterial lumen. Mediators of inflammation are not only instrumental in the formation of plaque, but may also be involved in the rapid progression of atheromatous lesions, leading to plaque fissuring and intraluminal thrombosis. Several gene products including lipoprotein receptors, adhesion molecules, growth factors, cytokines, matrix metalloproteinases and proteins associated with cell death are also involved (2-3). This suggests that proteins play an essential role in the formation and progression of plaque.The use ofproteomic-based strategies, examining the protein content of cells or tissues, offers a useful approach for the study of plaque proteins (4). Proteins obtained from fragments of tissue with atherosclerotic lesions were recently studied in vitro (5-6), revealing an increase in transport proteins (Apo AIV, haptoglobin, transferrin, actin), signal transduction proteins (serum amyloid protein P) and a decrease in shock protein 27. The only proteins detected directly in coronary and carotid

Proteomic analysis and protein carbonylation profile in trained and untrained rat muscles

“…This type of gel is obtained by averaging the positions, shapes and optical densities of the spots in a given set of gels [43,44]. Fig.…”

Overview of Current Proteomic Approaches for Discovery of Vascular Biomarkers of Atherosclerosis

“…In this report, authors identified cathepsin D as a potential target for therapeutical treatment of atherosclerosis. Terzuoli et al, 2007;Porcelli et al, 2010) IMAC, immobilized metal affinity chromatography. ↑, increase.…”

“…In this way, they identified 19 proteins with differential distribution between ICA and CCA segments. To overcome the problem of plaque heterogeneity, Terzuoli et al proposed a method for selecting proteins exclusive to plaque by constructing a reference synthetic gel (Terzuoli et al, 2007;Porcelli et al, 2010). This gel, obtained by averaging the positions, shapes and optical densities of spots in 2DE maps from 10 carotid plaque samples was compared with an equivalent synthetic gel constructed using 10 plasma samples from the same carotid surgery patients.…”