In vivo quantitative proteomics: the SILAC mouse

Methods Mol Biol. 2012;757:435-50. doi: 10.1007/978-1-61779-166-6_25.

Abstract

Mass spectrometry-based proteomics is a field that has been quickly developing, enabling increasingly giving in-depth characterization of the proteomes of cells and tissues. Current technology allows identifying thousands of proteins in a single experiment. Stable isotope labeling with amino acid in cell culture (SILAC) was originally developed for high accuracy quantitative proteomic studies in cell lines. We have shown that SILAC can be extended to in vivo animal model by fully labeling C57BL/6 mice with (13)C(6)-Lysine (Lys6). We used SILAC mice technology to map quantitative proteomic changes in mice lacking the expression of β1 integrin, β-Parvin, or the integrin tail-binding protein Kindlin-3. This approach confirmed the absence of the proteins and revealed a role of Kindlin-3 in red blood cells. Here we describe a practical method to generate and maintain a colony of SILAC mice and optimal strategies to perform in vivo quantitative proteomic experiments.

MeSH terms

  • Amino Acids / metabolism*
  • Animals
  • Carbon Isotopes
  • Female
  • Isotope Labeling / methods*
  • Mass Spectrometry*
  • Mice
  • Mice, Inbred C57BL
  • Protein Processing, Post-Translational
  • Proteomics*

Substances

  • Amino Acids
  • Carbon Isotopes