OFFgel-based multidimensional LC-MS/MS approach to the cataloguing of the human platelet proteome for an interactomic profile

Shibu Krishnan, Marco Gaspari, Anna Della Corte, Patrizia Bianchi, Marilena Crescente, Chiara Cerletti, Daniele Torella, Ciro Indolfi, Giovanni de Gaetano, Maria Benedetta Donati, Domenico Rotilio, Giovanni Cuda

Research output: Contribution to journalArticlepeer-review

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

Original languageEnglish
Pages (from-to)686-695
Number of pages10
JournalElectrophoresis
Volume32
Issue number6-7
DOIs
Publication statusPublished - Mar 2011

Keywords

  • Interactomics
  • Nano-LC-MS/MS
  • OFFgel electrophoresis
  • Platelet proteome
  • Shotgun proteomics

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry

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