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Novel Online Three-Dimensional Separation Expands the Detectable Functional Landscape of Cellular Phosphoproteome
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- Authors
- Issue Date
- 2022-09
- Publisher
- American Chemical Society
- Citation
- Analytical Chemistry, Vol.94 No.35, pp.12185-12195
- Abstract
- Protein phosphorylation is a prevalent post-translational modification that regulates essentially every aspect of cellular processes. Currently, liquid chromatography-tandem mass spectrometry (LC-MS/MS) with an extensive offline sample fractionation and a phosphopeptide enrichment method is a best practice for deep phosphoproteome profiling, but balancing throughput and profiling depth remains a practical challenge. We present an online three-dimensional separation method for ultradeep phosphoproteome profiling that combines an online two-dimensional liquid chromatography separation and an additional gas-phase separation. This method identified over 100,000 phosphopeptides (>60,000 phosphosites) in HeLa cells during 1.5 days of data acquisition, and the largest HeLa cell phosphoproteome significantly expanded the detectable functional landscape of cellular phosphoproteome.
- ISSN
- 0003-2700
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