Turnover of the human proteome: determination of protein intracellular stability by dynamic SILAC

J Proteome Res. 2009 Jan;8(1):104-12. doi: 10.1021/pr800641v.


The proteome of any system is a dynamic entity, such that the intracellular concentration of a protein is dictated by the relative rates of synthesis and degradation. In this work, we have analyzed time-dependent changes in the incorporation of a stable amino acid resolved precursor, a protocol we refer to as "dynamic SILAC", using 1-D gel separation followed by in-gel digestion and LC-MS/MS analyses to profile the intracellular stability of almost 600 proteins from human A549 adenocarcinoma cells, requiring multiple measures of the extent of labeling with stable isotope labeled amino acids in a classic label-chase experiment. As turnover rates are acquired, a profile can be built up that allows exploration of the 'dynamic proteome' and of putative features that predispose a protein to a high or a low rate of turnover. Moreover, measurement of the turnover rate of individual components of supramolecular complexes provides a unique insight in processes of protein complex assembly and turnover.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line, Tumor
  • Databases, Protein
  • Electrophoresis, Polyacrylamide Gel
  • Gene Expression Regulation, Neoplastic
  • Humans
  • Kinetics
  • Mass Spectrometry / methods*
  • Peptide Mapping / methods
  • Protein Processing, Post-Translational
  • Proteins / chemistry*
  • Proteome
  • Proteomics / methods*
  • Ribosomes / chemistry
  • Stochastic Processes
  • Time Factors


  • Proteins
  • Proteome