Synthetic phosphopeptides: from spike-in standards to affinity tools for protein-protein interaction studies
Synthetic isotope labeled phosphopeptides are valuable tools for the quantification and validation of phosphoproteome data. Here, we report that the same set of phosphopeptides, which are used as spike-in standards, can be successfully applied for identification of stimulus specific protein-protein...
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| Main Authors: | , , , , |
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| Format: | Article (Journal) |
| Language: | English |
| Published: |
2019
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| In: |
Analytical biochemistry
Year: 2018, Volume: 568, Pages: 73-77 |
| ISSN: | 1096-0309 |
| DOI: | 10.1016/j.ab.2018.12.018 |
| Online Access: | Verlag, Volltext: https://doi.org/10.1016/j.ab.2018.12.018 Verlag, Volltext: http://www.sciencedirect.com/science/article/pii/S0003269718312739 
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| Author Notes: | Martin Winter, Ramona Mayer, Uwe Warnken, Jürgen Debus, Amir Abdollahi, Martina Schnölzer |
| Summary: | Synthetic isotope labeled phosphopeptides are valuable tools for the quantification and validation of phosphoproteome data. Here, we report that the same set of phosphopeptides, which are used as spike-in standards, can be successfully applied for identification of stimulus specific protein-protein interactions mediated by the respective phosphorylation sites. As a proof-of-concept, binding of two γH2AX (pS139) phosphosite specific interaction partners, MDC1 and 53BP1, was confirmed and elevated binding affinity was revealed in response to ionizing radiation. Our strategy is generally applicable and enables multiplexed validation and functional analysis of phosphorylation sites offering great potential for the follow-up of phosphoproteome studies. |
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| Item Description: | Available online 28 December 2018 Gesehen am 19.09.2019 |
| Physical Description: | Online Resource |
| ISSN: | 1096-0309 |
| DOI: | 10.1016/j.ab.2018.12.018 |