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Updated: Jul 15, 2026

In-vitro Reconstitution of Bacterial Ubiquitination and VCP/p97-mediated Elimination
Published on: January 2, 2026
Detailed structural insights into the p97-Npl4-Ufd1 interface.
Rivka L Isaacson1, Valerie E Pye, Peter Simpson
1Division of Molecular Biosciences, Imperial College London, Biochemistry Building, South Kensington, London SW7 2AZ, United Kingdom.
The AAA ATPase p97 uses the UN adaptor complex (Ufd1-Npl4) for cellular functions. Structural analysis reveals how Npl4
Area of Science:
- Biochemistry
- Structural Biology
- Molecular Cell Biology
Background:
- The AAA ATPase p97 is a versatile protein involved in various cellular processes.
- Adaptor proteins, such as the UN complex (Ufd1-Npl4), mediate p97's diverse functions, particularly in the ubiquitin pathway.
- Structural understanding of p97's interaction with its adaptors is limited.
Purpose of the Study:
- To elucidate the structural basis of p97 recognition by the UN adaptor complex.
- To characterize the Npl4 ubiquitin-like domain (UBD) structure and its binding interface with p97.
- To investigate the interaction dynamics within the full-length p97-UN complex.
Main Methods:
- Nuclear Magnetic Resonance (NMR) spectroscopy, including chemical shift perturbation (CSP) analysis.
- Solution structure determination of the Npl4 UBD.
- Structural modeling of the p97 N-Npl4 UBD complex.
Main Results:
- The Npl4 UBD adopts a beta-grasp fold with a 3(10) helical insert.
- NMR data identified the binding surfaces between the p97 N domain and the Npl4 UBD.
- The interaction mode is conserved in the full-length p97-UN complex and differs subtly from p97-p47 interactions.
Conclusions:
- This study provides crucial structural insights into the p97-UN adaptor recognition mechanism.
- The findings highlight the specificity determinants in p97-adaptor interactions.
- Understanding these interactions is vital for deciphering p97's role in cellular regulation and disease.
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