Differential Impact of p97 Inhibitor CB-5083 on Protein Dynamics in Nucleus, Cytoplasm, and Membrane Compartments

Human p97/VCP is a vital AAA ATPase (ATPase associated with diverse cellular activity) plays critical roles in autophagy, endosomal trafficking, and the ubiquitin-proteasome system. Mutations in p97 cause inclusion body myopathy associated with paget’s disease of bone and frontotemporal dementia. P97 is overexpressed in several cancers. The selective active site inhibitor CB-5083 targets p97’s D2 domain and shows potential as an anti-cancer therapeutic, though it has faced clinical setbacks due to off-target effects. To investigate the protein levels changed of HL60 cells (acute myeloid leukemia cell line) by CB-5083 in cytoplasmic, nuclear, and insoluble membrane compartments, we employed fractionation and label-free proteomic analysis. Results reveal distinct compartment-specific protein regulation, providing insight into CB5083’s cellular mechanisms and potential for more targeted therapeutic applications.