Novel phosphorylation substrates reveal a spatially regulated role for AAK1 in cell migration - SILAC proteome profiling to determine the impact of acute AAK1/BMP2K inhibition

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 and its paralog BMP2K, as their only known role until now has been in endocytosis. Using SILAC labelling for whole cell proteome and phosphoproteome profiling of htert RPE cells pharmacological perturbation of these kinases, we demonstrate that acute inhibition of AAK1 and BMP2K significantly disrupts the abundance and phosphorylation of focal adhesion proteins. This indicates that AAK1 and BMP2K are involved in maintaining focal adhesions in addition to their role in endocytosis.