Tools
Tools
Caffrey, Brian 
ORCID: https://orcid.org/0000-0001-5040-4247, Zhu, Xing, Berezuk, Alison, Tuttle, Katharine, Chittori, Sagar and Subramaniam, Sriram
(2021)
AAA+ ATPase p97/VCP mutants and inhibitor binding disrupt inter-domain coupling and subsequent allosteric activation.
Journal of Biological Chemistry, 297 (4).
ISSN 0021-9258
Preview |
PDF (1-s2.0-S0021925821009893-main)
- Published Version
Available under License Creative Commons Attribution. Download (3MB) | Preview |
Abstract
The human AAA+ ATPase p97, also known as valosincontaining protein, a potential target for cancer therapeutics, plays a vital role in the clearing of misfolded proteins. p97 dysfunction is also known to play a crucial role in several neurodegenerative disorders, such as MultiSystem Proteinopathy 1 (MSP-1) and Familial Amyotrophic Lateral Sclerosis (ALS). However, the structural basis of its role in such diseases remains elusive. Here, we present cryo-EM structural analyses of four disease mutants p97R155H, p97R191Q, p97A232E, p97D592N, as well as p97E470D, implicated in resistance to the drug CB-5083, a potent p97 inhibitor. Our cryo-EM structures demonstrate that these mutations affect nucleotide-driven allosteric activation across the three principal p97 domains (N, D1, and D2) by predominantly interfering with either (1) the coupling between the D1 and N-terminal domains (p97R155H and p97R191Q), (2) the interprotomer interactions (p97A232E), or (3) the coupling between D1 and D2 nucleotide domains (p97D592N, p97E470D). We also show that binding of the competitive inhibitor, CB-5083, to the D2 domain prevents conformational changes similar to those seen for mutations that affect coupling between the D1 and D2 domains. Our studies enable tracing of the path of allosteric activation across p97 and establish a common mechanistic link between active site inhibition and defects in allosteric activation by diseasecausing mutations and have potential implications for the design of novel allosteric compounds that can modulate p97 function.
| Item Type: | Article |
|---|---|
| Additional Information: | Data availability: The density maps and refined atomic models have been deposited in the Electron Microscopy Data Bank with accession numbers EMD-24518, 24519, 24522, 24523, 24524, 24525, 24526, 24528, 24529, 24530, 24531, and 24532 and in the Protein Data Bank with matching accession numbers of PDB-7RL6, 7RL7, 7RL9, 7RLA, 7RLB, 7RLC, 7RLD, 7RLF, 7RLG, 7RLH, 7RLI, and 7RLJ respectively, for R155H-ADP, R155H-ATPγS, R191Q-ADP, R191Q-ATPγS, A232E-ADP, A232E-ATPγS, E470D-ADP, E470D-ATPγS, D592N-ADP, D592N-ATPγS mutant p97, and for p97 bound to CB-5083-ADP and CB-5083-ATPγS. All other data are available from the corresponding authors upon request. |
| Uncontrolled Keywords: | biochemistry,molecular biology,cell biology,sdg 3 - good health and well-being ,/dk/atira/pure/subjectarea/asjc/1300/1303 |
| Faculty \ School: | Faculty of Science > School of Biological Sciences |
| Related URLs: | |
| Depositing User: | LivePure Connector |
| Date Deposited: | 10 Jul 2026 15:54 |
| Last Modified: | 12 Jul 2026 23:02 |
| URI: | https://ueaeprints.uea.ac.uk/id/eprint/103824 |
| DOI: | 10.1016/j.jbc.2021.101187 |
Downloads
Downloads per month over past year
Actions (login required)
 |
View Item |