Fragment screening targeting Ebola virus nucleoprotein C-terminal domain identifies lead candidates

David J. Aceti, Hamza Ahmed, William M. Westler, Chao Wu, Hesam Dashti, Marco Tonelli, Hamid Eghbalnia, Gaya K. Amarasinghe, John L. Markley

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

The Ebola Virus is a causative agent of viral hemorrhagic fever outbreaks and a potential global health risk. The outbreak in West Africa (2013–2016) led to 11,000+ deaths and 30,000+ Ebola infected individuals. The current outbreak in the Democratic Republic of Congo (DRC) with 3000+ confirmed cases and 2000+ deaths attributed to Ebola virus infections provides a reminder that innovative countermeasures are still needed. Ebola virus encodes 7 open reading frames (ORFs). Of these, the nucleocapsid protein (eNP) encoded by the first ORF plays many significant roles, including a role in viral RNA synthesis. Here we describe efforts to target the C-terminal domain of eNP (eNP-CTD) that contains highly conserved residues 641–739 as a pan-Ebola antiviral target. Interactions of eNP-CTD with Ebola Viral Protein 30 (eVP30) and Viral Protein 40 (eVP40) have been shown to be crucial for viral RNA synthesis, virion formation, and virion transport. We used nuclear magnetic response (NMR)-based methods to screened the eNP-CTD against a fragment library. Perturbations of 1D 1H NMR spectra identified of 48 of the 439 compounds screened as potential eNP CTD interactors. Subsequent analysis of these compounds to measure chemical shift perturbations in 2D 1H,15N NMR spectra of 15N-labeled protein identified six with low millimolar affinities. All six perturbed an area consisting mainly of residues at or near the extreme C-terminus that we named “Site 1” while three other sites were perturbed by other compounds. Our findings here demonstrate the potential utility of eNP as a target, several fragment hits, and provide an experimental pipeline to validate viral-viral interactions as potential panfiloviral inhibitor targets.

Original languageEnglish
Article number104822
JournalAntiviral Research
Volume180
DOIs
StatePublished - Aug 2020

Keywords

  • Drug development
  • Ebola virus
  • Fragment screening
  • Nucleoprotein C-terminal domain

Fingerprint

Dive into the research topics of 'Fragment screening targeting Ebola virus nucleoprotein C-terminal domain identifies lead candidates'. Together they form a unique fingerprint.

Cite this