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
ROR:	https://ror.org/023b0x485
DOI:	http://doi.org/10.25358/openscience-9391
Version:	Published version
Publication type:	Zeitschriftenaufsatz
License:	CC BY
Information on rights of use:	https://creativecommons.org/licenses/by/4.0/
Journal:	Journal of molecular medicine 101
Pages or article number:	855 867
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