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Updated: Jun 24, 2025
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A Mass Spectrometry-Based Approach to Identify Phosphoprotein Phosphatases and their Interactors
Published on: April 29, 2022
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Integrated Platform for Large-Scale Quantitative Profiling of Phosphotyrosine Signaling Complexes Based on
Wei Liu1, Xuantang Wang1, Hailong Yu1
1Department of Chemistry, Institutes of Biomedical Sciences, Fudan University, Shanghai 200438, China.
Analytical Chemistry
|June 5, 2024
Summary
Researchers developed a new platform to study phosphotyrosine (pTyr) protein complexes. This method quantifies pTyr complexes, revealing their composition and dynamics in cells, advancing signaling pathway research.
Area of Science:
- Proteomics
- Cell Signaling
- Biochemistry
Background:
- Phosphotyrosine (pTyr) modified proteins are crucial in cell signaling but challenging to study due to their dynamic nature and low abundance.
- Understanding pTyr protein complexes is vital for deciphering complex cellular processes and disease mechanisms.
Purpose of the Study:
- To develop an integrated, complex-centric platform for large-scale quantitative profiling of pTyr signaling complexes.
- To establish a novel computational approach for quantifying protein complexes based on their elution profiles.
Main Methods:
- Development of a trifunctional probe (SH2-S) for specific isolation of intact pTyr protein complexes.
- Application of cofractionation/mass spectrometry (CoFrac-MS) with ion exchange chromatography and data-independent acquisition mass spectrometry.
- Creation of a complex-centric algorithm for quantitative analysis of protein complex elution curves.
Main Results:
- Successfully quantified 216 putative pTyr protein complexes using the developed platform and algorithm.
- Identified and screened 21 regulated pTyr protein complexes associated with epidermal growth factor signaling.
- Generated a quantitative landscape of pTyr protein complex composition in HeLa cells.
Conclusions:
- The study presents a comprehensive framework for the intricate examination of pTyr protein complexes.
- The developed platform enables quantitative profiling, offering new insights into pTyr signaling networks.
- This work provides the first quantitative map of pTyr protein complex composition in human cells.
