Characterization of an Aptamer Targeting the p47/p97 Complex for Therapeutic and Diagnostic Applications in Neurodegenerative Disorders and Cancer

p97 is an essential AAA+ ATPase involved in many cellular processes, and its dysregulation is implicated in cancer and neurodegenerative diseases such as ALS. Direct inhibition of p97 is challenging due to its central role in cellular pathways. To achieve pathway-specific modulation, this work targets the p97 cofactor p47, which directs p97 to membrane fusion processes. Using capture-SELEX, an aptamer with high affinity and specificity for p47 was identified. ITC revealed micromolar binding affinity, while NMR spectroscopy localized the interaction to the SEP domain. Chemical shift perturbation and fluorescence assays mapped the protein binding interface and key nucleobases on the aptamer. Importantly, the aptamer retained binding to p47 in the presence of p97, demonstrating compatibility with the native complex. Functional assays probed the impact of aptamer binding on p47-mediated p97 activity. Together, these results provide a framework for understanding aptamer recognition of p47 and establish another pathway for specific modulators of p97.

Grantor dc:publisher

University of Guelph

Author dc:creator

• Kim, Peter

Advisors dc:contributor.advisor

• Huang, Rui
• O'Flaherty, Derek

• aptamer
• NMR
• p47

Statement dc:rights

• Attribution-NonCommercial-ShareAlike 4.0 International

Licence dc:rights.uri

• http://creativecommons.org/licenses/by-nc-sa/4.0/

Language dc:language.iso

en

Handle dc:identifier.uri

https://hdl.handle.net/10214/29618

OAI record oai:identifier

https://atrium.lib.uoguelph.ca/server/oai/request?verb=GetRecord&metadataPrefix=dim&identifier=oai%3Aatrium.lib.uoguelph.ca%3A10214%2F29618