title: OFFgel-based multidimensional LC-MS/MS approach to the cataloguing of the human platelet proteome for an interactomic profile.
abstract: The proteome of quiescent human platelets was analyzed by a shotgun proteomics approach consisting of enzymatic digestion, peptide separation based on isoelectric point by the use of OFFgel fractionation and, finally, RP nanoscale chromatography coupled to MS/MS detection (nano-LC-MS/MS). OFFgel fractionation in the first dimension was effective in providing an additional dimension of separation, orthogonal to RP nano-LC, thus generating an off-line multidimensional separation platform that proved to be robust and easy to set up. The analysis identified 1373 proteins with high confidence (false discovery rate<0.25%). The core set of 1373 human platelet proteins was investigated by Ingenuity Pathway Analysis software from which ten canonical pathways and eight networks have been validated, to suggest that platelets behave either as inflammatory or immune cells, and plasma membrane and cytoskeleton proteins play a fundamental role in their function. Moreover, toxicity pathway in agreement with network analysis, supports the concept that platelet life span is governed by an apoptotic mechanism.
authors.affiliation: Laboratory of Proteomics and Mass Spectrometry and Department of Experimental and Clinical Medicine, "Magna Graecia" University of Catanzaro, Catanzaro, Italy.
authors.forename: Shibu
authors.initials: S
authors.lastname: Krishnan
authors.forename: Marco
authors.initials: M
authors.lastname: Gaspari
authors.forename: Anna
authors.initials: A
authors.lastname: Della Corte
authors.forename: Patrizia
authors.initials: P
authors.lastname: Bianchi
authors.forename: Marilena
authors.initials: M
authors.lastname: Crescente
authors.forename: Chiara
authors.initials: C
authors.lastname: Cerletti
authors.forename: Daniele
authors.initials: D
authors.lastname: Torella
authors.forename: Ciro
authors.initials: C
authors.lastname: Indolfi
authors.forename: Giovanni
authors.initials: G
authors.lastname: de Gaetano
authors.forename: Maria Benedetta
authors.initials: MB
authors.lastname: Donati
authors.forename: Domenico
authors.initials: D
authors.lastname: Rotilio
authors.forename: Giovanni
authors.initials: G
authors.lastname: Cuda
mesh_terms: D001792:Blood Platelets / Q000737:chemistry*; D001798:Blood Proteins / Q000032:analysis* / Q000378:metabolism; D056148:Chromatography, Reverse-Phase / Q000379:methods*; D030562:Databases, Protein; D006801:Humans; D007525:Isoelectric Focusing; D010446:Peptide Fragments / Q000032:analysis / Q000097:blood; D010449:Peptide Mapping; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results; D012984:Software; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014357:Trypsin / Q000378:metabolism
chemical_list: D001798:Blood Proteins; D010446:Peptide Fragments; D020543:Proteome; D014357:Trypsin