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Catch me if you can: Mass spectrometry‐based phosphoproteomics and quantification strategies
Abstract
Phosphorylation of proteins is one of the most prominent PTMs and for instance a key regulator of signal transduction. In order to improve our understanding of cellular phosphorylation events, considerable effort has been devoted to improving the analysis of phosphorylation by MS‐based proteomics. Different enrichment strategies for phosphorylated peptides/proteins, such as immunoaffinity chromatography (IMAC) or titanium dioxide, have been established and constantly optimized for subsequent MS analysis. Concurrently, specific MS techniques were developed for more confident identification and phosphorylation site localization. In addition, more attention is paid to the LC‐MS instrumentation to avoid premature loss of phosphorylated peptides within the analytical system. Despite major advances in all of these fields, the analysis of phosphopeptides still remains far from being routine in proteomics. However, to reveal cellular regulation by phosphorylation events, not only qualitative information about the phosphorylation status of proteins but also, in particular, quantitative information about distinct changes in phosphorylation patterns upon specific stimulation is mandatory. Thus, yielded insights are of outstanding importance for the emerging field of systems biology. In this review, we will give an insight into the historical development of phosphoproteome analysis and discuss its recent progress particularly regarding phosphopeptide quantification and assessment of phosphorylation stoichiometry.
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“Thus, despite the high confidence of localization and sensitivity of such advanced phospho-enrichment MS techniques, a complete determination of the phosphorylation profile of a given protein still remains a challenge (37).”
From Identification of a Key Lysine Residue in Heat Shock Protein 90 Required for Azole and Echinocandin Resistance in Aspergillus fumigatus (2014) · cites this paper as [37]
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Research works printed but not linked: 1
- Hemmings H. D., 1996, Protein kinase and phosphatase inhibitors, Neuromethods, 30, 121
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Universal and Confident Phosphorylation Site Localization Using phosphoRS
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Quantitative mass spectrometry in proteomics: critical review update from 2007 to the present
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The next level of complexity: Crosstalk of posttranslational modifications
Venne, A. S., Kollipara, L., & Zahedi, R. P. (2013) · PROTEOMICS · doi:10.1002/pmic.201300344 · Cited by 331
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Kalli, A., Smith, G. T., Sweredoski, M. J., & Hess, S. Y. (2013) · Journal of Proteome Research · doi:10.1021/pr3011588 · Cited by 179
Metadata
- Authors
- Eyrich, B., Sickmann, A., & Zahedi, R. P.
- Affiliations
- Leibniz Institute for Analytical Sciences - ISAS (DE)
- Corresponding author
- Zahedi, René Peiman
- Venue
- PROTEOMICS
- Publisher
- Wiley
- Published
- 2011-01-11
- Volume
- 11
- Issue
- 4
- Pages
- 554-570
- ISSN
- 1615-9853 (ISSN-L)
- 1615-9861
- OpenAlex
- W2109482992
- Cited by
- 104
- References
- 178
- Citations by year
- 2026: 2 · 2025: 2 · 2024: 3 · 2023: 2 · 2022: 1 · 2021: 3 · 2020: 1 · 2019: 3 · 2018: 2 · 2017: 4 · 2016: 7 · 2015: 12 · 2014: 10 · 2013: 18 · 2012: 21
- FWCI
- 1.62
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- PROTEOMICS (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Ministerium für Innovation, Wissenschaft, Forschung und Technologie des Landes Nordrhein-Westfalen
- Bundesministerium für Bildung und Forschung (31P5800)
- Topics
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Physical Sciences › Chemistry › Spectroscopy › Mass Spectrometry Techniques and Applications
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Tyrosine Phosphatases
- Keywords
- phosphoproteomics, protein phosphorylation, phosphopeptide quantification, phosphorylation stoichiometry, phosphopeptide enrichment, immunoaffinity chromatography, titanium dioxide enrichment, phosphorylation site localization, signal transduction, systems biology
- MeSH terms
- Phosphoproteins, Mass Spectrometry, Proteomics
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