Dataset Information

Metabolic labelling and LC-MS/MS-based identification of IL-1α-induced secreted proteomes from epithelial cells in the presence or absence of serum

ABSTRACT: The unbiased identification of cytokine-induced, secreted proteins from cells cultured in serum-containing medium is challenging. Here, we describe an experimental and bioinformatics workflow to label IL-1α-regulated proteins in living cells with the methionine analogue L-homopropargylglycine (L-HPG), purify and identify them by means of a CLICK-chemistry based biotinylation step followed by nanoHPLC-MS/MS. A side-by-side comparison of enriched proteins and their ontologies to serum-free conditions demonstrates the sensitivity and specificity of this approach to study the inducible secreted proteomes of epithelial cells.

INSTRUMENT(S): timsTOF Pro

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Permanent Cell Line Cell, Epithelial Cell, Colon

SUBMITTER: Axel Weber

LAB HEAD: Michael Kracht

PROVIDER: PXD040974 | Pride | 2023-03-21

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	220622_DR_251_Sf_10_Slot2-16_1_11263.d.zip	Other
	220622_DR_251_Sf_10a_Slot2-16_1_11294.d.zip	Other
	220622_DR_251_Sf_11_Slot2-17_1_11264.d.zip	Other
	220622_DR_251_Sf_11a_Slot2-17_1_11295.d.zip	Other
	220622_DR_251_Sf_12_Slot2-18_1_11265.d.zip	Other

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Metabolic labeling and LC-MS/MS-based identification of interleukin-1α-induced secreted proteomes from epithelial cells in the presence or absence of serum.

Priester Jasmin J Meier-Soelch Johanna J Weiser Hendrik H Heylmann Daniel D Weber Axel A Linne Uwe U Kracht Michael M

STAR protocols 20230331 2

The unbiased identification of cytokine-induced, secreted proteins from cells cultured in serum-containing medium is challenging. Here, we describe an experimental and bioinformatics workflow to label interleukin-1α-regulated proteins in living cells with the methionine analogue L-homopropargylglycine. We detail their purification and identification by means of CLICK-chemistry-based biotinylation followed by nanoHPLC-MS/MS. A side-by-side comparison of enriched proteins and their ontologies to s ...[more]

PMID: 37004159

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