Trans-SILAC: sorting out the non-cell-autonomous proteome

Nat Methods. 2010 Nov;7(11):923-7. doi: 10.1038/nmeth.1513. Epub 2010 Oct 10.

Abstract

Non-cell-autonomous proteins are incorporated into cells that form tight contacts or are invaded by bacteria, but identifying the full repertoire of transferred proteins has been a challenge. Here we introduce a quantitative proteomics approach to sort out non-cell-autonomous proteins synthesized by other cells or intracellular pathogens. Our approach combines stable-isotope labeling of amino acids in cell culture (SILAC), high-purity cell sorting and bioinformatics analysis to identify the repertoire of relevant non-cell-autonomous proteins. This 'trans-SILAC' method allowed us to discover many proteins transferred from human B to natural killer cells and to measure biosynthesis rates of Salmonella enterica proteins in infected human cells. Trans-SILAC should be a useful method to examine protein exchange between different cells of multicellular organisms or pathogen and host.

MeSH terms

  • B7-2 Antigen / metabolism
  • Bacterial Proteins / biosynthesis
  • Cell Communication
  • Cells, Cultured
  • Humans
  • Isotope Labeling / methods*
  • Protein Biosynthesis
  • Protein Transport*
  • Proteome*

Substances

  • B7-2 Antigen
  • Bacterial Proteins
  • CD86 protein, human
  • Proteome