Novel phosphorylation substrates reveal a spatially regulated role for AAK1 in cell migration - AAK1 C-terminal interactome profiling – batch 2,3

NAKs (Numb-associated kinases) are a relatively unexplored kinase family that interacts with the endocytic machinery. Among NAK members, AAK1 has been linked to disorders such as neuropathic pain, schizophrenia, and Alzheimer's disease. Our goal is to better understand the cellular pathways regulated by AAK1 as its only known role until now has been in endocytosis. Using a synthetic peptide identical to the C-terminus of AAK1, we observed an interaction with both the clathrin heavy chain and a subset of proteins containing the PDZ domain, including the newly identified AAK1 substrate, PDLIM5. This finding is significant as it reveals that AAK1 and PDLIM5 interact not only through the kinase domain of AAK1 and PDLIM5's consensus motif surrounding the threonine 290 (T290) but also through the PDZ domain of PDLIM5 binding the LIDL motif at the C-terminus of AAK1.