Detection of the Ubiquitinome in Cells Undergoing Oncogene-Induced Senescence

Methods Mol Biol. 2017:1534:127-137. doi: 10.1007/978-1-4939-6670-7_12.

Abstract

Senescent cells exhibit dramatic changes in protein post-translational modifications. Here, we describe a method, stable isotope labeling with amino acids in cell culture (SILAC) coupled to liquid chromatography tandem mass spectrometry (LC-MS/MS), to identify changes in the ubiquitinome in cells that have undergone oncogene-induced senescence.

Keywords: Liquid chromatography tandem mass spectrometry (LC-MS/MS); Post-translational modification; Senescence; Stable isotope labeling with amino acids in cell culture (SILAC); Ubiquitin; Ubiquitin-proteosome pathway; Ubiquitinome.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Cell Line
  • Cellular Senescence* / genetics
  • Chromatography, Affinity
  • Chromatography, Liquid / methods
  • Fibroblasts / metabolism
  • Genes, ras
  • Humans
  • Isotope Labeling
  • Oncogenes / genetics*
  • Protein Processing, Post-Translational
  • Proteome*
  • Proteomics* / methods
  • Tandem Mass Spectrometry / methods
  • Ubiquitins / isolation & purification
  • Ubiquitins / metabolism*

Substances

  • Proteome
  • Ubiquitins