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Updated: Oct 16, 2025

Phospho Flow Cytometry with Fluorescent Cell Barcoding for Single Cell Signaling Analysis and Biomarker Discovery
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Phosphoproteome profiling uncovers a key role for CDKs in TNF signaling

Maria C Tanzer1, Isabell Bludau2, Che A Stafford3

  • 1Department of Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Martinsried, 82152, Germany. tanzer@biochem.mpg.de.

Nature Communications
|October 19, 2021
PubMed

Insights

This study used systems proteomics to map tumor necrosis factor (TNF) signaling pathways. Findings reveal key roles for protein phosphorylation and kinase activity in regulating cellular responses to TNF, aiding targeted therapy development.

Area of Science:

  • Immunology
  • Cell Biology
  • Proteomics

Background:

  • Tumor necrosis factor (TNF) is a crucial cytokine in inflammation and immunity.
  • Targeting TNF is effective for inflammatory diseases but causes side effects due to its pleiotropic nature.
  • Understanding TNF's complex signaling is vital for developing safer therapies.

Purpose of the Study:

  • To perform a systems-level proteomic analysis of TNF signaling.
  • To identify functional modules of protein phosphorylation and their upstream kinases.
  • To uncover mechanisms regulating TNF-induced cell death and cytokine production.

Main Methods:

  • Time-course phosphoproteomics experiments.
  • Subcellular localization studies.
  • Kinase inhibitor analysis.
  • Spatial proteomics.

Main Results:

  • Identified functional modules of protein phosphorylation in TNF signaling.
  • Assigned most regulated phosphorylation events to specific upstream kinases.
  • Revealed phosphorylation-dependent protein translocations upon TNF stimulation.
  • Uncovered a role for cyclin-dependent kinase activity in promoting cytokine production and preventing cell death.

Conclusions:

  • Systems-level proteomic analysis provides insights into TNF signaling complexity.
  • Understanding phosphorylation dynamics aids in developing targeted TNF-based therapies.
  • The TNF signaling resource is available at http://tnfviewer.biochem.mpg.de/ .

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