The need for scalable methods for functional annotation of phosphotyrosine sites has led the Olsen group, Novo Nordisk Foundation Center for Protein Research in Copenhagen, to develop a new mass spectrometry-based workflow for high-throughput analyses. In a recent study, they implemented this new workflow used to describe how cancer mutations close to tyrosine phosphorylation sites in the EGF receptor rewire signaling pathways by switching protein interactors.
To be able to decipher phosphotyrosine signaling in tissue samples the Olsen Group and colleagues have combined DIA with short LC gradients. The workflow used consists of an automated sample preparation method and analysis of samples run on our Evosep One LC system coupled to a Thermo Scientific™ Q Exactive™ HF-X mass spectrometer with fast MS/MS scanning capabilities. This method furthermore provides the possibility for a scalable methodology, meaning 1200 pulldown experiments can be analyzed in just 20 days of LC-MS instrument time.
Recent data using Evosep One
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Oncogenic Mutations Rewire Signaling Pathways by Switching Protein Recruitment to Phosphotyrosine Sites was published by Olsen and colleagues in Cell on October 3. https://www.cell.com/cell/fulltext/S0092-8674(19)31014-1