SILAC-IodoTMT for Assessment of the Cellular Proteome and Its Redox Status

Methods Mol Biol. 2023:2603:259-268. doi: 10.1007/978-1-0716-2863-8_21.

Abstract

Stable isotope labeling by amino acids in cell culture (SILAC) and iodoacetyl tandem mass tag (iodoTMT) are well-implemented mass spectrometry-based approaches for quantification of proteins and for site-mapping of cysteine modification. We describe here a combination of SILAC and iodoTMT to assess ongoing changes in the global proteome and cysteine modification levels using liquid chromatography separation coupled with high-resolution mass spectrometry (LC-MS/MS).

Keywords: Cysteine; Global proteome; IodoTMT; Liquid chromatography; Mass spectrometry; Redox; SILAC.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatography, Liquid / methods
  • Cysteine / metabolism
  • Isotope Labeling / methods
  • Oxidation-Reduction
  • Proteome* / metabolism
  • Proteomics* / methods
  • Tandem Mass Spectrometry / methods

Substances

  • Proteome
  • Cysteine