Aims: Recently, the apolipoprotein composition of the major lipoprotein classes as VLDL, LDL, and HDL in physiological conditions has been studied by means of proteomic approaches. The aim of this work was to characterize the apolipoprotein pattern in atherosclerotic patients by means of two-dimensional electrophoresis and mass spectrometry. Methods: For this purpose we purified plasma VLDL, LDL, and HDL from patients undergoing carotid endarterectomy and from healthy normolipidemic subjects by single isopycnic density gradient ultracentrifugation. Samples were loaded into 4-7 IPG strips and isoelectric focused. Proteins were then reduced and alkylated before SDS-PAGE. After staining and image analysis, spots of interest were excised and protein identified by peptide mass fingerprinting after tryptic digestion. Results: We identified 21 spots corresponding to about 96% of 52 protein spots detected in VLDL, 22 spots corresponding to about 92% of 43 spots in LDL and 20 corresponding to about 96% of 60 spots in HDL. By image analysis we found many differences in the relative abundance of some identified VLDL and LDL apolipoproteins between patients and healthy subjects. Conversely, HDL showed quite similar apolipoprotein profiles. Conclusions: In summary, we have set up a method to compare 2DE apolipoproteins profiles of plasma VLDL, LDL and HDL fractions. This method seems to be suitable to detect potential changes associated to the atherosclerotic process, and it could provide insight into the development of novel diagnostic tools and/or future therapeutic agents. This work was supported by “Fondazione Banco di Sardegna” (Sassari, Italy) grant

Apolipoprotein profiles in atherosclerotic disease / Formato, Marilena; Lepedda, Antonio Junior; Zinellu, E; Cigliano, A; Piredda, F; Bacciu, Pp; Guarino, A; Spirito, R.. - (2011). (Intervento presentato al convegno 79th EAS Congress tenutosi a Gothenburg, Sweden nel June 26-29, 2011).

Apolipoprotein profiles in atherosclerotic disease

FORMATO, Marilena;LEPEDDA, Antonio Junior;Cigliano A;
2011-01-01

Abstract

Aims: Recently, the apolipoprotein composition of the major lipoprotein classes as VLDL, LDL, and HDL in physiological conditions has been studied by means of proteomic approaches. The aim of this work was to characterize the apolipoprotein pattern in atherosclerotic patients by means of two-dimensional electrophoresis and mass spectrometry. Methods: For this purpose we purified plasma VLDL, LDL, and HDL from patients undergoing carotid endarterectomy and from healthy normolipidemic subjects by single isopycnic density gradient ultracentrifugation. Samples were loaded into 4-7 IPG strips and isoelectric focused. Proteins were then reduced and alkylated before SDS-PAGE. After staining and image analysis, spots of interest were excised and protein identified by peptide mass fingerprinting after tryptic digestion. Results: We identified 21 spots corresponding to about 96% of 52 protein spots detected in VLDL, 22 spots corresponding to about 92% of 43 spots in LDL and 20 corresponding to about 96% of 60 spots in HDL. By image analysis we found many differences in the relative abundance of some identified VLDL and LDL apolipoproteins between patients and healthy subjects. Conversely, HDL showed quite similar apolipoprotein profiles. Conclusions: In summary, we have set up a method to compare 2DE apolipoproteins profiles of plasma VLDL, LDL and HDL fractions. This method seems to be suitable to detect potential changes associated to the atherosclerotic process, and it could provide insight into the development of novel diagnostic tools and/or future therapeutic agents. This work was supported by “Fondazione Banco di Sardegna” (Sassari, Italy) grant
2011
Apolipoprotein profiles in atherosclerotic disease / Formato, Marilena; Lepedda, Antonio Junior; Zinellu, E; Cigliano, A; Piredda, F; Bacciu, Pp; Guarino, A; Spirito, R.. - (2011). (Intervento presentato al convegno 79th EAS Congress tenutosi a Gothenburg, Sweden nel June 26-29, 2011).
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11388/54512
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