Stable isotope dynamic labeling of secretomes (SIDLS) identifies authentic secretory proteins released by cancer and stromal cells

Dean E Hammond, J. Dinesh Kumar, Lorna Raymond, Deborah M. Simpson, Robert J. Beynon, Graham J. Dockray, Andrea Varro

Research output: Contribution to journalArticlepeer-review

11 Citations (Scopus)

Abstract

Analysis of secretomes critically underpins the capacity to understand the mechanisms determining interactions between cells and between cells and their environment. In the context of cancer cell micro-environments, the relevant interactions are recognised to be an important determinant of tumor progression. Global proteomic analyses of secretomes are often performed at a single time point and frequently identify both classical secreted proteins (possessing an N-terminal signal sequence), as well as many intracellular proteins, the release of which is of uncertain biological significance. Here, we describe a mass spectrometry-based method for stable isotope dynamic labeling of secretomes (SIDLS) that, by dynamic SILAC, discriminates the secretion kinetics of classical secretory proteins and intracellular proteins released from cancer and stromal cells in culture. SIDLS is a robust classifier of the different cellular origins of proteins within the secretome and should be broadly applicable to non-proliferating cells and cells grown in short term culture.

Original languageEnglish
JournalMolecular and Cellular Proteomics
VolumeFirst Online
Early online date18 Jun 2018
DOIs
Publication statusE-pub ahead of print - 18 Jun 2018

Keywords

  • Short term labelling
  • Dynamic labelling
  • Cancer biology
  • Cell secretion
  • Exocytosis
  • Secretome
  • SILAC

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