[Frontiers in Bioscience E2, 796-804, June 1, 2010]

[Frontiers in Bioscience E2, 796-804, June 1, 2010]

Proteomics reveals high levels of vitamin D binding protein in myocardial infarction

Cosimo Gasparri^1,2, Antonio Curcio^1,2, Daniele Torella^1,2, Marco Gaspari³, Vittoria Celi⁴, Francesco Salituri¹, Duino Boncompagni¹, Michele Torella⁵, Elio Gulletta⁴, Giovanni Cuda³, Ciro Indolfi^1,2

1Division of Cardiology,"Magna Graecia" University, Catanzaro, Italy, ²Laboratory of Molecular and Cellular Cardiology,"Magna Graecia" University, Catanzaro, Italy, ³Laboratory of Proteomics and Mass Spectrometry,"Magna Graecia" University, Catanzaro, Italy, ⁴Department of Experimental and Clinical Medicine,"Magna Graecia" University, Catanzaro, Italy, ⁵Division of Cardiac Surgery, Second University of Naples, Naples, Italy

TABLE OF CONTENTS

1. Abstract
2. Introduction
3. Materials and Methods: 3.1. Sample preparation; 3.2. ICAT assay; 3.3. Protein sequencing and identification; 3.4. Western blotting analysis on whole serum; 3.5. Immunoblotting analysis in thrombus extracts; 3.6. Platelet Aggregation Study; 3.7. Coagulation assays; 3.8. Data Analyses and Statistics
4. Results: 4.1. Proteomic analysis revealed novel serum fingerprints; 4.2. VDB protein is increased in thrombi extracts; 4.3. VDB protein administration decreased platelet aggregation; 4.4. VDB protein administration affects coagulation cascade at different levels 5. Discussion
6. Acknowledgments
7. References

1. ABSTRACT

The pathogenic mechanisms underlying the disease processes in cardiovascular disease are likely to involve significant alterations in myocardial gene and protein expression. Proteomics analysis can define new protein and peptide changes associated with myocardial infarction (MI). The aim of the present study was to analyze serum proteome of patients with ST-Elevation MI (STEMI). Serum samples were collected from STEMI patients (age 65.0+/-10.3) at 5.3+/-2.7 hours after the onset of typical chest pain and before initiating standard therapy. Ten age- and sex-matched donors were used as controls. The samples were albumin- and IgG-depleted. Isotope-coded affinity tag method was employed to label cysteine residues and liquid chromatography-Tandem Mass Spectrometry analysis was performed to measure the labelled proteins. Our proteomic approach identified increased levels of vitamin D-binding protein precursor (VDB) in the serum from STEMI patients compared to control donors. Western blot analysis confirmed the increase in VDB protein in STEMI patients. Moreover, fresh thrombotic plaques, obtained during primary angioplasty, showed high expression of VDB protein. Finally, VDB protein reduced the aggregation rate and prolonged coagulation time.