Targeted Covalent Inhibitors Allosterically Deactivate the DEDDh Lassa Fever Virus NP Exonuclease from Alternative Distal Sites

Kuan Wei Huang, Jing Wen Chen, Tzu Yu Hua, Yu Yu Chu, Tsai Yuan Chiu, Jung Yu Liu, Chun I. Tu, Kai Cheng Hsu, Ya Ting Kao, Jhih Wei Chu, Yu Yuan Hsiao

Research output: Contribution to journalArticlepeer-review

Abstract

For using targeted covalent inhibitors (TCIs) as anticancer and antiviral drugs, we establish that the model compounds PCMPS (p-chloromercuriphenyl sulfate) and PCMB (p-chloromercuribenzoate) are inhibitors of the DEDDh family of exonucleases. The underlying mechanism is analyzed by X-ray crystallography, activity/nucleic acid-binding assays, and all-atom molecular dynamics (MD) simulations. The first TCI-complexed structures of a DEDDh enzyme, the Lassa fever virus NP exonuclease (NPexo), are resolved to elucidate that the Cys409 binding site is away from the active site and the RNA-binding lid. The NPexo C409A structures indicate Cys461 as the alternative distal site for obstructing the equally active mutant. All-atom MD simulations of the wild type and mutant NPexos in explicit solvent uncover an allosteric inhibition mechanism that the local perturbation induced by PCMPS sulfonate propagates to impact the RNA-binding lid conformation. Binding assay studies confirm that PCMPS does affect the RNA binding of NPexo. The predicted relative potency between PCMPS and PCMB is also in line with experiments. The structural data and inhibition mechanism established in this work provide an important molecular basis for the drug development of TCIs.

Original languageEnglish
Pages (from-to)2315-2327
Number of pages13
JournalJournal of the American Chemical Society
Volume1
Issue number12
DOIs
Publication statusPublished - Dec 27 2021

Keywords

  • DEDDh exonuclease
  • inhibitor
  • molecular dynamics
  • NP exonuclease
  • organomercurial

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

Fingerprint

Dive into the research topics of 'Targeted Covalent Inhibitors Allosterically Deactivate the DEDDh Lassa Fever Virus NP Exonuclease from Alternative Distal Sites'. Together they form a unique fingerprint.

Cite this