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Andreas‐David Brunner, Marvin Thielert, Catherine G. Vasilopoulou, Constantin Ammar, Fabian Coscia, Andreas Mund, Ole B. Hørning, Nicolai Bache, Amalia Apalategui, Markus Lubeck and 7 more
Single-cell technologies are revolutionizing biology but are today mainly limited to imaging and deep sequencing 1–3 . 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 4,5 . Chemical labeling-based single-cell approaches introduce hundreds of cells into the MS, but direct analysis of single cells has not yet reached the necessary sensitivity, robustness and quantitative accuracy to answer biological questions 6,7 . 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 ten-fold improved sensitivity. We accurately 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 such as CDK2NA, the E2 ubiquitin ligase UBE2S, DNA topoisomerases TOP2A/B and the chromatin regulator HMGA1. 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 volatile. This emphasizes substantial regulation of translation and sets the stage for its elucidation at the single cell level. Our technology can readily be applied to ultra-high sensitivity analyses of tissue material 8 , posttranslational modifications and small molecule studies to gain unprecedented insights into cellular heterogeneity in health and disease.
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Brunner, A., Thielert, M., Vasilopoulou, C. G., Ammar, C., Coscia, F., Mund, A., Hørning, 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. (2022). Ultra‐high sensitivity mass spectrometry quantifies single‐cell proteome changes upon perturbation. Molecular Systems Biology, 18(3), e10798. https://doi.org/10.15252/msb.202110798
Brunner, Andreas‐David, et al. "Ultra‐high sensitivity mass spectrometry quantifies single‐cell proteome changes upon perturbation." Molecular Systems Biology, vol. 18, no. 3, 2022, pp. e10798, https://doi.org/10.15252/msb.202110798.
Brunner, Andreas‐David, Marvin Thielert, Catherine G. Vasilopoulou, Constantin Ammar, Fabian Coscia, Andreas Mund, Ole B. Hørning, et al. "Ultra‐high sensitivity mass spectrometry quantifies single‐cell proteome changes upon perturbation." Molecular Systems Biology 18, no. 3 (2022): e10798. https://doi.org/10.15252/msb.202110798.
Brunner A., Thielert M., Vasilopoulou C. G. [и др.] Ultra‐high sensitivity mass spectrometry quantifies single‐cell proteome changes upon perturbation // Molecular Systems Biology. — 2022. — Vol. 18, № 3. — P. e10798. — URL: https://doi.org/10.15252/msb.202110798
@article{brunner2022ultrahigh,
title = {{Ultra‐high sensitivity mass spectrometry quantifies single‐cell proteome changes upon perturbation}},
author = {Andreas‐David Brunner and Marvin Thielert and Catherine G. Vasilopoulou and Constantin Ammar and Fabian Coscia and Andreas Mund and Ole B. Hørning and Nicolai Bache and Amalia Apalategui and Markus Lubeck and Sabrina Richter and David S. Fischer and Oliver Raether and Melvin A. Park and Florian Meier and Fabian Joachim Theis and Matthias Mann},
journal = {Molecular Systems Biology},
year = {2022},
volume = {18},
number = {3},
pages = {e10798},
doi = {10.15252/msb.202110798},
url = {https://doi.org/10.15252/msb.202110798}
}TY - JOUR AU - Brunner, Andreas‐David AU - Thielert, Marvin AU - Vasilopoulou, Catherine G. AU - Ammar, Constantin AU - Coscia, Fabian AU - Mund, Andreas AU - Hørning, Ole B. AU - Bache, Nicolai AU - Apalategui, Amalia AU - Lubeck, Markus AU - Richter, Sabrina AU - Fischer, David S. AU - Raether, Oliver AU - Park, Melvin A. AU - Meier, Florian AU - Theis, Fabian Joachim AU - Mann, Matthias TI - Ultra‐high sensitivity mass spectrometry quantifies single‐cell proteome changes upon perturbation JO - Molecular Systems Biology VL - 18 IS - 3 SP - e10798 PY - 2022 DA - 2022/02/28 DO - 10.15252/msb.202110798 UR - https://doi.org/10.15252/msb.202110798 ER -
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