Deep mutational scanning of Met kinase

MET kinase is critical in the initiation and regulation of several signaling pathways, which can be disrupted through disease mutations that alter MET activity. Cancer-associated mutations have been identified throughout the intracellular domain, including point mutations within the kinase domain and exon14 skipping. Comprehensively mapping all possible kinase mutations within the context of a wild type or exon14 skipped intracellular domain would not only provide a reference for potential novel and rare activating mutations, but also reveal how specific mutations may alter the activation equilibrium in each context. Here, we perform a parallel deep mutational scan of the MET kinase domain in a wild type and exon14 skipped background to gain insight into MET regulation and activating mutations. With this approach, we have identified unique MET mutational sensitivities, clinically relevant and previously undescribed activating mutations, and provided a framework for comparative, and targeted analysis.