Updated project metadata. Here we use high-resolution mass spectrometry (MS) to characterise the magnitude and complexity of global tyrosine phosphorylation PDAC. We profiled the endogenous tyrosine phosphorylation-based signaling in 36 cell lines from two cell line populations, quantifying around 2,000 tyrosine phosphorylation sites. This approach facilitated consistent segregation of PDAC cell lines into three subtypes with distinct signalling profiles, using both their global tyrosine phosphorylation patterns and a Random Forest derived 8 phosphorylation site signature. We extracted segregation-driving phosphosites, and subsequent pathway and network analysis to characterise the identified subtypes.