Advancements in Global Phosphoproteomics Profiling: Overcoming Challenges in Sensitivity and Quantification

Area of Science:

• Biochemistry
• Molecular Biology
• Proteomics

Background:

• Protein phosphorylation is a key post-genomic modification driving cellular activities.
• Understanding signaling cascades necessitates precise identification and quantification of phosphorylation dynamics.
• Phosphoproteomic technologies map dynamic phosphorylation landscapes, aiding exploration of cell-specific networks.

Purpose of the Study:

• To provide an overview of phosphoproteomics basics, challenges, and technological evolution.
• To summarize state-of-the-art global and quantitative phosphoproteomics methodologies.
• To highlight innovations enabling deep profiling from low-input samples.

Main Methods:

• Overview of technological evolution in phosphoproteomics.
• Focus on miniaturized sample preparation for micro/nano-scale and single-cell profiling.
• Application of data-independent acquisition mass spectrometry (DIA-MS) for enhanced coverage.
• Utilization of quantitative phosphoproteomic pipelines for deep mapping.

Main Results:

• Recent advances enable comprehensive mapping of the dynamic phosphoproteomic landscape.
• Innovations include miniaturized sample prep, DIA-MS, and quantitative pipelines.
• Evolving technologies facilitate deep profiling from low-input samples.

Conclusions:

• Phosphoproteomics offers new avenues for exploring cell function and clinical phenotypes.
• Technological advancements address challenges in nanoscale and single-cell profiling.
• Future outlooks focus on improving phosphoproteomic technologies for biomedical applications.