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

Hammond, DE ORCID: 0000-0002-6326-8739, Kumar, JD, Raymond, L, Simpson, DM, Beynon, RJ, Dockray, GJ and Varro, A (2018) Stable isotope dynamic labeling of secretomes (SIDLs) identifies authentic secretory proteins released by cancer and stromal cells. Molecular and Cellular Proteomics, 17 (9). pp. 1837-1849. ISSN 1535-9476

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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 recognized 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 nonproliferating cells and cells grown in short term culture.

Item Type:	Article
Uncontrolled Keywords:	Cell Line, Tumor; Intracellular Space; Stromal Cells; Humans; Neoplasms; Proteome; Reproducibility of Results; Isotope Labeling; Signal Transduction; Kinetics; Time Factors; Gene Ontology; Cell Line, Tumor; Gene Ontology; Humans; Intracellular Space; Isotope Labeling; Kinetics; Neoplasms; Proteome; Reproducibility of Results; Signal Transduction; Stromal Cells; Time Factors; 3101 Biochemistry and Cell Biology; 31 Biological Sciences; Biotechnology; Cancer; 1.1 Normal biological development and functioning; 2.1 Biological and endogenous factors; Cancer; Cell Line, Tumor; Gene Ontology; Humans; Intracellular Space; Isotope Labeling; Kinetics; Neoplasms; Proteome; Reproducibility of Results; Signal Transduction; Stromal Cells; Time Factors; Biochemistry & Molecular Biology
Subjects:	Q Science > QH Natural history > QH301 Biology R Medicine > RC Internal medicine > RC0254 Neoplasms. Tumors. Oncology (including Cancer)
Divisions:	Pharmacy and Biomolecular Sciences
Publisher:	Elsevier
Date of acceptance:	18 June 2018
Date of first compliant Open Access:	7 July 2026
Date Deposited:	07 Jul 2026 12:28
Last Modified:	07 Jul 2026 12:28
DOI or ID number:	10.1074/mcp.TIR117.000516
URI:	https://researchonline.ljmu.ac.uk/id/eprint/28973

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