Dataset Information

Stable isotope dynamic labelling of secretomes: Strataclean Linearity C4PR_LIV

ABSTRACT: We improve on currently-available resources by describing a mass spectrometry (MS)-based strategy using stable isotope dynamic labelling of secretomes (SIDLS) that discriminates between authentic secretory proteins and intracellular proteins within the secretome of cultured cells. By monitoring the rate of incorporation of labelled amino acids into newly synthesised proteins as they appear in the media, we can differentiate those proteins that have been destined for secretion, and exhibit rapid labelling, from those with low rates of labelling or low turnover relative to the growth rate of the cells which is a feature of intracellular proteins. Part of the wet lab protocol for our analysis of secretomes, is the use of the resin, Strataclean (Agilent). In this Supplementary dataset to our main paper (also in PRIDE/ProteomeXchange), we characterise the linearity of capture of secreted proteins using Stracalean resin, by label-free quantitative mass spectrometry. We mix cell-coditioned culture media with 'virgin' media in different ratios and analayse the capture capacity of equal quantitites of StrataClean reagent.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture, Fibroblast

DISEASE(S): Esophagus Adenocarcinoma

SUBMITTER: Dean Hammond

LAB HEAD: Dean Hammond

PROVIDER: PXD009838 | Pride | 2019-10-08

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	DS_070318_0_100_2h_1uL.raw	Raw
	DS_070318_100_0_2h_1uL.raw	Raw
	DS_070318_20_80_2h_1uL.raw	Raw
	DS_070318_50_50_2h_1uL.raw	Raw
	DS_070318_80_20_2h_1uL.raw	Raw

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Publications

Stable Isotope Dynamic Labeling of Secretomes (SIDLS) Identifies Authentic Secretory Proteins Released by Cancer and Stromal Cells.

Hammond Dean E DE Kumar J Dinesh JD Raymond Lorna L Simpson Deborah M DM Beynon Robert J RJ Dockray Graham J GJ Varro Andrea A

Molecular & cellular proteomics : MCP 20180618 9

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 wel ...[more]

PMID: 29915148

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