Identifying Functionally Relevant Proteins and Phosphorylation Sites During T Cell Activation Using an Integrated Thermal Shift Assay

In this study, we sought to identify how protein thermal stability integrates with changes in cell signaling and protein abundance. We developed a pipeline to investigate changes in protein abundance and thermal stabilite by combining the protein abundance and thermal shift measurements into the same TMT-plex in the Integrated Thermal Shift Assay (ITSA). Additionally, to elucidate how protein signaling corresponds to changes in thermal stability we also examined protein phosphorylation in the Phosphorylation Integrated Thermal Shift Assay (PITSA).