Related Experiment Video
Updated: Jun 16, 2025
10:17
A Mass Spectrometry-Based Approach to Identify Phosphoprotein Phosphatases and their Interactors
Published on: April 29, 2022
2.4K
Fast and deep phosphoproteome analysis with the Orbitrap Astral mass spectrometer
Noah M Lancaster1,2, Pavel Sinitcyn3, Patrick Forny4
1Department of Chemistry, University of Wisconsin-Madison, Madison, WI, USA.
Nature Communications
|August 15, 2024
Summary
A new mass spectrometry method enables rapid, deep analysis of protein phosphorylation sites in humans and mice. This technology advances phosphoproteomics, revealing thousands of phosphorylation sites for biological discovery.
Area of Science:
- Biochemistry
- Proteomics
- Cell Biology
Background:
- Protein phosphorylation is crucial for cellular signal transduction and cell processes.
- Detecting and quantifying protein phosphorylation remains a significant analytical challenge.
Purpose of the Study:
- To introduce a novel mass spectrometry approach for rapid and deep phosphoproteome analysis.
- To benchmark the new technology against existing state-of-the-art platforms.
- To generate a comprehensive mouse phosphoproteome atlas and explore phosphorylation contexts.
Main Methods:
- Utilized a novel mass spectrometer (Orbitrap Astral) combined with data-independent acquisition (DIA).
- Analyzed human and mouse phosphoproteomes, including synthetic peptide standards and EGF-stimulated HeLa cells.
- Generated a multi-tissue phosphoproteome atlas of the mouse.
Main Results:
- Mapped approximately 30,000 unique human phosphorylation sites in 30 minutes.
- Detected 81,120 unique phosphorylation sites in the mouse across 12 hours of measurement.
- Examined sequence, structural, and kinase specificity contexts of identified phosphorylation sites.
Conclusions:
- The Orbitrap Astral MS platform coupled with DIA provides rapid and deep phosphoproteome analysis.
- This technology significantly advances the scale and speed of phosphoproteomics research.
- The generated datasets offer a valuable resource for exploring phosphorylation events in various biological contexts, including mitochondrial and brain biology.
