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Signal Transduction Reaction Monitoring Deciphers Site-Specific PI3K-mTOR/MAPK Pathway Dynamics in Oncogene-Induced

Erik L de Graaf1,2, Joanna Kaplon3, Shabaz Mohammed1,2

  • 1†Biomolecular Mass Spectrometry and Proteomics, Utrecht Institute for Pharmaceutical Sciences and Bijvoet Center for Biomolecular Research, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands.

Journal of Proteome Research
|May 27, 2015
PubMed

Insights

We developed a sensitive method to track signaling pathway changes in mammals using targeted proteomics. This approach reveals new phosphorylation events in cancer-related pathways, aiding drug development.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cell Signaling

Background:

  • Signal transduction pathways are crucial for cellular functions.
  • Understanding phosphorylation dynamics is key to deciphering cellular responses.
  • Existing methods for pathway analysis can be complex and lack sensitivity.

Purpose of the Study:

  • To develop a straightforward and sensitive strategy for monitoring signal transduction pathway dynamics in mammalian systems.
  • To analyze phosphorylation dynamics in the PI3K-mTOR and MAPK signaling networks.
  • To identify previously uncharacterized phosphorylation changes in oncogene-induced senescence.

Main Methods:

  • Utilized targeted quantitative proteomics combined with highly selective phosphopeptide enrichment.
  • Developed a single-step enrichment followed by a single targeted proteomics experiment.
  • Circumvented the need for labeling and immune purification.

Main Results:

  • Achieved sensitive monitoring of phosphorylation dynamics in the PI3K-mTOR and MAPK signaling networks.
  • Identified uncharacterized phosphorylation changes in oncogene-induced senescence.
  • Demonstrated the utility of the approach in understanding biological phenotypes and pharmacological interventions.

Conclusions:

  • The developed strategy offers a sensitive and comprehensive approach to monitor signal transduction pathway dynamics.
  • This method facilitates the analysis of specific phosphorylation nodes across signaling pathways.
  • The findings highlight the importance of comprehensive pathway analysis in understanding cellular processes and disease states.
Keywords:
ERK1/2MAPKPI3KRPS6SRMTi4+-IMACmTORnetwork biologypathwayphosphoproteomicssignal transduction

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