Extending SILAC to proteomics of plant cell lines

Plant Cell. 2011 May;23(5):1701-5. doi: 10.1105/tpc.110.082016. Epub 2011 May 3.

Abstract

Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) is a widespread method for metabolic labeling of cells and tissues in quantitative proteomics; however, incomplete incorporation of the label has so far restricted its wider use in plants. Here, we argue that differential labeling by two different versions of the labeled amino acids renders SILAC fully applicable to dark-grown plant cell lines. By comparing Arabidopsis thaliana cell cultures labeled with two versions of heavy Lys (Lys-4 and Lys-8), we show that this simple modification of the SILAC protocol enables similar quantitation accuracy, precision, and reproducibility as conventional SILAC in animal cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Amino Acids / chemistry*
  • Amino Acids / metabolism
  • Arabidopsis / cytology
  • Arabidopsis / metabolism*
  • Arabidopsis Proteins / metabolism*
  • Cell Culture Techniques
  • Cell Line
  • Humans
  • Isotope Labeling / methods*
  • Lysine / chemistry
  • Lysine / metabolism
  • Mass Spectrometry
  • Molecular Sequence Data
  • Proteomics / methods*
  • Reproducibility of Results
  • Time Factors

Substances

  • Amino Acids
  • Arabidopsis Proteins
  • Lysine