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Brunner, A.D.* ; Thielert, M.* ; Vasilopoulou, C.G.* ; Ammar, C.* ; Coscia, F.* ; Mund, A.* ; Hoerning, O.B.* ; Bache, N.* ; Apalategui, A.* ; Lubeck, M.* ; Richter, S. ; Fischer, D.S. ; Raether, O.* ; Park, M.A.* ; Meier, F.* ; Theis, F.J. ; Mann, M.*

Ultra-high sensitivity mass spectrometry quantifies single-cell proteome changes upon perturbation.

Mol. Syst. Biol. 18:e10798 (2022)
Publ. Version/Full Text Research data DOI
Open Access Gold
Creative Commons Lizenzvertrag
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.
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Publication type Article: Journal article
Document type Scientific Article
Keywords Drug Perturbation ; Low-flow Lc-ms ; Proteomics At Single-cell Resolution ; Single-cell Heterogeneity ; Systems Biology
ISSN (print) / ISBN 1744-4292
e-ISSN 1744-4292
Quellenangaben Volume: 18, Issue: 3, Pages: , Article Number: e10798 Supplement: ,
Publisher EMBO Press
Reviewing status Peer reviewed
Grants Graduate School of Quantitative Biosciences Munich
Marie Curie individual fellowship
Deutsches Forschungszentrum für Gesundheit und Umwelt, Helmholtz Zentrum München (German Research Center for Environmental Health, Helmholtz Zentrum München)
Novo Nordisk Fonden (NNF)
Max-Planck-Gesellschaft (MPG)