Related Experiment Video
Updated: Feb 24, 2026
12:23
Phosphopeptide Enrichment Coupled with Label-free Quantitative Mass Spectrometry to Investigate the Phosphoproteome in Prostate Cancer
Published on: August 2, 2018
12.8K
Sensitive, Robust, and Cost-Effective Approach for Tyrosine Phosphoproteome Analysis
Mingming Dong1,2, Yangyang Bian1,3, Yan Wang1,2
1CAS Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences (CAS) , Dalian 116023, China.
Analytical Chemistry
|August 11, 2017
Summary
A new SH2 superbinder method enhances tyrosine phosphoproteome analysis, identifying 41% more phosphotyrosine (pTyr) peptides. This sensitive and cost-effective approach improves the study of cellular signaling pathways.
Area of Science:
- Biochemistry
- Proteomics
- Cellular Signaling
Background:
- Tyrosine phosphorylation (pTyr) is crucial for cell signaling but challenging to analyze at the proteome level.
- Conventional antibody-based methods for pTyr enrichment lack sensitivity, reproducibility, and are costly.
Purpose of the Study:
- To develop and validate an efficient workflow for sensitive tyrosine phosphoproteome analysis using an SH2 domain-derived superbinder.
- To overcome the limitations of traditional antibody-based enrichment methods for pTyr peptides.
Main Methods:
- Utilized an SH2 superbinder for specific enrichment of phosphotyrosine (pTyr) peptides.
- Applied the new workflow to analyze tyrosine phosphoproteome in various biological samples, including cell lines and tissue.
Main Results:
- Identified 41% more pTyr peptides compared to previous methods.
- Detected over 1800 high-confidence pTyr sites from 2 mg of pervanadate-treated Jurkat T cells.
- Identified 343 high-confidence pTyr sites from 5 mg of unstimulated Jurkat cells, a 31% increase over antibody methods.
- Characterized 197 high-confidence pTyr sites from mouse skeletal muscle tissue.
Conclusions:
- The SH2 superbinder-based workflow offers a sensitive, robust, and cost-effective approach for tyrosine phosphoproteome analysis.
- This method significantly enhances the identification of pTyr sites, even in low-abundance samples.
- The approach has broad applications for studying the role of tyrosine phosphorylation in health and disease.
