Ultra-high sensitivity mass spectrometry quantifies single-cell proteome changes upon perturbation.
drug perturbation
low-flow LC-MS
proteomics at single-cell resolution
single-cell heterogeneity
systems biology
Journal
Molecular systems biology
ISSN: 1744-4292
Titre abrégé: Mol Syst Biol
Pays: England
ID NLM: 101235389
Informations de publication
Date de publication:
03 2022
03 2022
Historique:
revised:
06
02
2022
received:
08
11
2021
accepted:
08
02
2022
entrez:
28
2
2022
pubmed:
1
3
2022
medline:
3
5
2022
Statut:
ppublish
Résumé
Single-cell technologies are revolutionizing biology but are today mainly limited to imaging and deep sequencing. However, proteins are the main drivers of cellular function and in-depth characterization of individual cells by mass spectrometry (MS)-based proteomics would thus be highly valuable and complementary. Here, we develop a robust workflow combining miniaturized sample preparation, very low flow-rate chromatography, and a novel trapped ion mobility mass spectrometer, resulting in a more than 10-fold improved sensitivity. We precisely and robustly quantify proteomes and their changes in single, FACS-isolated cells. Arresting cells at defined stages of the cell cycle by drug treatment retrieves expected key regulators. Furthermore, it highlights potential novel ones and allows cell phase prediction. Comparing the variability in more than 430 single-cell proteomes to transcriptome data revealed a stable-core proteome despite perturbation, while the transcriptome appears stochastic. Our technology can readily be applied to ultra-high sensitivity analyses of tissue material, posttranslational modifications, and small molecule studies from small cell counts to gain unprecedented insights into cellular heterogeneity in health and disease.
Identifiants
pubmed: 35226415
doi: 10.15252/msb.202110798
pmc: PMC8884154
doi:
Substances chimiques
Proteome
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e10798Informations de copyright
© 2022 The Authors Published under the terms of the CC BY 4.0 license.
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