Cryo-EM of the disease mutant p97 bound with its cofactor p47
EMSL Project ID
51010
Abstract
Mutation in p97/VCP (valosin-containing protein), an ATPase associated with diverse cellular activities, has been implicated in IBMPFD/ALS, which currently does not have an effective medical intervention for the treatment. Particularly, the R155H p97 mutant is responsible for at least half of the disease prevalence. Thus, the mechanistic view into the p97 action is essential to understand the pathophysiological cause of the related disease. A structural perspective in how p97 mutations dysregulate its functional role is critical to design small molecule or bio-drug therapeutics for treatments.Human p97 binds to p47 and VCIP135 to regulate membrane fusion in Golgi apparatus and endoplasmic reticulum via VCIP135 phosphorylation and its deubiquitination process on Syn5 protein. However, the structural basis of the interactions between these proteins as well as the disease mutants has not yet been determined. From existing knowledge and our preliminary studies, this proposed research will test the hypothesis that the R155H p97 mutant hinders the interaction of p97-p47 complex with VCIP135 and thereby disables the Syn5 deubiquitination.
To approach the goal, we will first characterize the binding of p97 with p47, which is the initial step in membrane fusion. The structure of the assembly will be determined using single-particle cryo-EM. The complex of the R155H mutant with p47 will be investigated in parallel to understand the disease role of the mutated residue. The knowledge of these protein-protein interactions will be improved in the understanding in molecular mechanism of p97-mediated membrane fusion.
Project Details
Start Date
2019-09-01
End Date
2020-03-20
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members