Understanding how the ATPase p97 functions at molecular level

Summary The AAA ATPase p97 is a ubiquitous, abundant and essential protein.

Project Details

It is involved in a multitude of pathways required for cell homeostasis and is involved in many pathways. In humans, p97 specificity is determined by interactions with more than 20 cofactors. Current models propose that p97 functions as a “segregase”, converting ATP energy into conformational changes in order to disassemble (segregate) proteins from protein-protein/DNA complexes or membranes. How conformational changes in p97 drive disassembly of complexes is unknown.

The general aim of the project is to understand at the molecular level how the AAA ATPase functions. Our approach is to determine 3D cryo-EM structures of p97 in different conformations, alone and in presence of co-factors, inhibitors and substrates. In parallel, we are developing a system in order to characterise using fluorescence spectroscopy (FRET) the dynamic properties of p97 and their modulation by disease-associated mutations, co-factors and inhibitors. This information will be combined with Molecular Dynamics to understand how p97 functions. There are multiple projects available. Projects involve mutagenesis, protein expression/purification, biochemical characterisation and single particle cryo-EM.

Impact of the project

Because of its central role in cell homeostasis, p97 is a target for cancer treatment. Besides, missense mutations in the p97 gene have been estimated to cause ~50% of multisystem proteinopathy (MSP), a dominantly inherited, pleiotropic, degenerative disorder of humans that can affect muscle, bone and/or the central nervous system. Drugs targeting p97 for the treatment of both cancer and MSP are at the early stages of development. Our studies will provide novel and unanticipated insight into the mechanism of action of the fundamental cellular enzyme p97, knowledge critical to develop pathways specific inhibitors.