Proteomics investigation of human platelets in healthy donors and cystic fibrosis patients by shotgun nUPLC-MS^Eand 2DE: a comparative study

Abstract

Platelets are of pathophysiological relevance in haemostasis, wound repair, inflammation and cardiovascular disease. We have shown that human platelets express a biologically active Cystic Fibrosis Transmembrane Conductance Regulator, which is dysfunctional in Cystic Fibrosis (CF) patients, and regulate platelet responses related to inflammation and its resolution. In order to further elucidate platelet involvement in CF inflammation, we pursued a comparative proteomic analysis of cells from healthy donors and CF patients, in association with a non-supervised comparative analysis of the Gene Ontology. Our results, showing changes in the integrin signalling in CF, support a pro-inflammatory profile of CF platelets.

Proteomic and ionomic profiling reveals significant alterations of protein expression and calcium homeostasis in cystic fibrosis cells

“…The high complementary potential of this approach to the 2DE protein analysis has been demonstrated in CF models. 17 Therefore, our study is expected to significantly integrate previous results obtained using 2DE-MS.…”

“…Identified proteins were normalized against P00924 entry (Enolase S. cerevisiae) in the quantitative analysis. 17,22 Gene ontology and pathway analysis Functional classification based on gene ontology (GO) terms related to biological processes and function, respectively, was performed using the web resource the Generic Gene Ontology Term Finder available at http//go.princeton.edu/cgi-bin/GOTerm Finder. 24 The GO Term Finder attempts to determine whether an observed level of annotation for a group of genes is significant within the context of annotation for all genes within the genome.…”

Exosomal clusterin, identified in the pericardial fluid, improves myocardial performance following MI through epicardial activation, enhanced arteriogenesis and reduced apoptosis

“…5B). To study the protein repertoire characteristic of PFC and PFMI-derived exosomes, we designed a Label Free Proteomics experiment by Data Independent Analysis based on a Shotgun discovery proteomics methodology [26]. This experimental setting allowed us to detect the most abundant proteins of each sample and indeed we identified 60 and 72 proteins in the plasma of control patients and patients with acute MI, respectively, and 69 and 65 in PFC and PFMI, respectively (Supplement Table III).…”

“…Continuum LC-MS data from four replicates experiments for each samples were processed for qualitative analysis using the software ProteinLynx Global Server v. 2.4 (PLGS, Waters) [26,28]. (Santa Cruz) followed by horseradish peroxidase-coupled secondary antibodies and developed by a chemiluminescence-based detection system (ECL, Amersham Pharmacia Biotech.…”

Copper exposure effects on yeast mitochondrial proteome

“…We have chosen to apply these two different technologies in order to gain the larger possible evaluation of the investigated protein repertories thus avoiding technique redundancy. Moreover, the partial overlapping of these two different experiments have been already reported in different systems and papers [22,26,39,40].…”