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Authors: Kirschner, Friederike
Arnold-Schild, Danielle
Leps, Christian
Łącki, Mateusz Krzysztof
Klein, Matthias
Chen, Yannic
Ludt, Annekathrin
Marini, Federico
Kücük, Can
Stein, Lara
Distler, Ute
Sielaff, Malte
Michna, Thomas
Riegel, Kristina
Rajalingam, Krishnaraj
Bopp, Tobias
Tenzer, Stefan
Schild, Hansjörg
Title: Modulation of cellular transcriptome and proteome composition by azidohomoalanine : implications on click chemistry–based secretome analysis
Online publication date: 18-Aug-2023
Year of first publication: 2023
Language: english
Abstract: The analysis of the secretome provides important information on proteins defining intercellular communication and the recruitment and behavior of cells in specific tissues. Especially in the context of tumors, secretome data can support decisions for diagnosis and therapy. The mass spectrometry–based analysis of cell-conditioned media is widely used for the unbiased characterization of cancer secretomes in vitro. Metabolic labeling using azide-containing amino acid analogs in combination with click chemistry facilitates this type of analysis in the presence of serum, preventing serum starvation-induced effects. The modified amino acid analogs, however, are less efficiently incorporated into newly synthesized proteins and may perturb protein folding. Combining transcriptome and proteome analysis, we elucidate in detail the effects of metabolic labeling with the methionine analog azidohomoalanine (AHA) on gene and protein expression. Our data reveal that 15–39% of the proteins detected in the secretome displayed changes in transcript and protein expression induced by AHA labeling. Gene Ontology (GO) analyses indicate that metabolic labeling using AHA leads to induction of cellular stress and apoptosis-related pathways and provide first insights on how this affects the composition of the secretome on a global scale.
DDC: 610 Medizin
610 Medical sciences
Institution: Johannes Gutenberg-Universität Mainz
Department: FB 04 Medizin
Place: Mainz
Version: Published version
Publication type: Zeitschriftenaufsatz
License: CC BY
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Journal: Journal of molecular medicine
Pages or article number: 855
Publisher: Springer
Publisher place: Berlin u.a.
Issue date: 2023
ISSN: 1432-1440
Publisher DOI: 10.1007/s00109-023-02333-4
Appears in collections:DFG-491381577-H

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