Abstract
Apolipoproteins are very heterogeneous protein family, implicated in plasma lipoprotein structural stabilization, lipid metabolism, inflammation, or immunity. Obtaining detailed information on apolipoprotein composition and structure may contribute to elucidating lipoprotein roles in atherogenesis and to developing new therapeutic strategies for the treatment of lipoprotein-associated disorders. This study aimed at developing a comprehensive method for characterizing the apolipoprotein component of plasma VLDL, LDL, and HDL fractions from patients undergoing carotid endarterectomy, by means of two-dimensional electrophoresis (2-DE) coupled with Mass Spectrometry analysis, useful for identifying potential markers of plaque presence and vulnerability. The adopted method allowed obtaining reproducible 2-DE maps of exchangeable apolipoproteins from VLDL, LDL, and HDL. Twenty-three protein isoforms were identified by peptide mass fingerprinting analysis. Differential proteomic analysis allowed for identifying increased levels of acute-phase serum amyloid A protein (AP SAA) in all lipoprotein fractions, especially in LDL from atherosclerotic patients. Results have been confirmed by western blotting analysis on each lipoprotein fraction using apo AI levels for data normalization. The higher levels of AP SAA found in patients suggest a role of LDL as AP SAA carrier into the subendothelial space of artery wall, where AP SAA accumulates and may exert noxious effects.
Highlights
Cardiovascular diseases are the leading cause of death and illness in developed countries, with atherosclerosis being the most important contributor
Mapping of Exchangeable Apolipoproteins from VLDL, LDL, and HDL Fractions. 2-DE combined with Mass Spectrometry was applied to identify VLDL, LDL, and HDL apolipoproteins from patients undergoing carotid endarterectomy and healthy normolipidemic subjects
Its levels were higher in all lipoprotein fractions, increasing up to fourteenfold in atherosclerotic patients LDL fraction
Summary
Cardiovascular diseases are the leading cause of death and illness in developed countries, with atherosclerosis being the most important contributor. Atherosclerosis is a chronic inflammatory condition that could turn into an acute clinical event due to plaque rupture and thrombosis [1]. Vascular inflammation plays a major role in the development of atherosclerosis and contributes to the acute onset of thrombotic complications [2]. The selective retention of circulating apolipoprotein B100 containing lipoproteins in the subendothelial space, by means of specific interactions with artery wall proteoglycans, is currently thought to be the leading event in atherogenesis [3, 4]. Lipoproteins are supramolecular complexes that deliver insoluble lipids from the tissues where they are synthesized to those that metabolize or store them They consist of hydrophobic molecules (core), triacylglycerol
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