Metabolic labeling and LC-MS/MS-based identification of interleukin-1α-induced secreted proteomes from epithelial cells in the presence or absence of serum.
Cell Biology
Cell Culture
Flow Cytometry/Mass Cytometry
Immunology
Molecular/Chemical Probes
Protein Biochemistry
Protein Expression and Purification
Proteomics
Journal
STAR protocols
ISSN: 2666-1667
Titre abrégé: STAR Protoc
Pays: United States
ID NLM: 101769501
Informations de publication
Date de publication:
31 Mar 2023
31 Mar 2023
Historique:
received:
29
11
2022
revised:
11
02
2023
accepted:
03
03
2023
entrez:
2
4
2023
pubmed:
3
4
2023
medline:
3
4
2023
Statut:
aheadofprint
Résumé
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 serum-free conditions demonstrates the sensitivity and specificity of this approach to study the inducible secreted proteomes of epithelial cells.
Identifiants
pubmed: 37004159
pii: S2666-1667(23)00153-3
doi: 10.1016/j.xpro.2023.102195
pmc: PMC10090805
pii:
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
102195Informations de copyright
Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of interests The authors declare no competing interests.
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