Tracking flaviviral protease conformational dynamics by tuning single-molecule nanopore tweezers.

ABSTRACT: The flaviviral NS2B/NS3 protease is a conserved enzyme required for flavivirus replication. Its highly dynamic conformation poses major challenges but also offers opportunities for antiviral inhibition. Here, we established a nanopore tweezers-based platform to monitor NS2B/NS3 conformational dynamics in real time. Molecular simulations coupled with single-channel current recording measurements revealed that the protease could be captured in the middle of the ClyA nanopore lumen, stabilized mainly by dynamic electrostatic interactions. We designed a new Salmonella typhi ClyA nanopore with enhanced nanopore/protease interaction that can resolve the open and closed states at the single-molecule level for the first time. We demonstrated that the tailored ClyA could track the conformational tr

SUBMITTER: Shorkey SA 

PROVIDER: S-EPMC11739873 | biostudies-literature | 2025 Jan

REPOSITORIES: biostudies-literature