Uncovering the molecular basis of kinase activity and substrate recognition with phospho-PCA.

ABSTRACT: Protein kinases relay information to various cellular processes, and their dysregulation underlies numerous human diseases. Despite their importance, our understanding of how kinase domains and their variants impact protein stability, catalytic activity, and substrate recognition is incomplete. In this work, we develop the phosphorylation protein complementation assay (phospho-PCA), which enables quantitative measurements of kinase-substrate interactions by coupling them to the growth of budding yeast, thereby enabling deep mutational scanning of kinase domains. When combined with deep mutational scans targeting folding stability and Bayesian modeling, phospho-PCA can disentangle the relative impact of mutations on kinase domain stability, catalytic activity, and substrate specificity. We

SUBMITTER: Yu C 

PROVIDER: S-EPMC12637564 | biostudies-literature | 2025 Nov

REPOSITORIES: biostudies-literature