Summary
With the appearance of a new generation of high-performance hybrid mass spectrometers, high accuracy (sub-parts-per-million) mass spectrometry is becoming increasingly available to a wider scientific commu nity. Here we discuss the advantages of such mass spectrometric instrumentation in the global analysis of protein phosphorylation. We describe a detailed workflow for fractionation and enrichment of phosphopep tides from digests of whole cell/tissue lysates by strong cation exchange and TiO 2 chromatography and their subsequent measurement on an LTQ-Orbitrap mass spectrometer under several acquisition regimes.
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Acknowledgments
The authors thank Prof. Matthias Mann and Dr. Chunaram Choudhary for critical reading of the manuscript and members of the Department for Proteomics and Signal Transduction at the MPI for Biochemistry for numerous discussions that led to the optimized protocols for phosphopeptide enrichment and MS analysis presented here.
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Olsen, J.V., Macek, B. (2009). High Accuracy Mass Spectrometry in Large-Scale Analysis of Protein Phosphorylation. In: Lipton, M.S., Paša-Tolic, L. (eds) Mass Spectrometry of Proteins and Peptides. Methods In Molecular Biology, vol 492. Humana Press. https://doi.org/10.1007/978-1-59745-493-3_7
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DOI: https://doi.org/10.1007/978-1-59745-493-3_7
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