Quantitative differential proteomics of cystic fibrosis cell models by SILAC (stable isotope labelling in cell culture)

Methods Mol Biol. 2011:742:213-25. doi: 10.1007/978-1-61779-120-8_13.

Abstract

Differential proteomics represents an enticing strategy to unmask the proteins involved in CF pathogenesis and to discover potential therapeutic targets and/or markers of disease progression. Quantitative proteomics is possible nowadays owing to the recent progress in protein labelling and/or in label-free approaches, combined to sensitive detection by mass spectrometry (MS). In this chapter, we present one strategy to perform differential quantitative proteomic studies on different cellular compartments of proliferating cell lines expressing wild-type (WT) CFTR and F508del-CFTR using stable isotope labelling in cell culture (SILAC).

MeSH terms

  • Cell Culture Techniques
  • Cell Line
  • Chromatography, High Pressure Liquid
  • Cystic Fibrosis / genetics
  • Cystic Fibrosis / metabolism
  • Cystic Fibrosis Transmembrane Conductance Regulator / chemistry*
  • Cystic Fibrosis Transmembrane Conductance Regulator / genetics*
  • Cystic Fibrosis Transmembrane Conductance Regulator / metabolism
  • Female
  • Humans
  • Isotope Labeling / methods*
  • Mass Spectrometry
  • Models, Biological
  • Mutation
  • Peptide Fragments / analysis*
  • Peptide Fragments / isolation & purification
  • Proteomics / methods*
  • Sequence Deletion
  • Trypsin / metabolism

Substances

  • Peptide Fragments
  • Cystic Fibrosis Transmembrane Conductance Regulator
  • Trypsin