A DARPin promotes faster onset of botulinum neurotoxin A1 action

Leka, Oneda; Wu, Yufan; Zanetti, Giulia; Furler, Sven; Reinberg, Thomas; Marinho, Joana; Schaefer, Jonas V; Plückthun, Andreas; Li, Xiaodan; Pirazzini, Marco; Kammerer, Richard A

doi:10.5167/uzh-252072

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Leka, O., Wu, Y., Zanetti, G., Furler, S., Reinberg, T., Marinho, J., Schaefer, J. V., Plückthun, A., Li, X., Pirazzini, M., & Kammerer, R. A. (2023). A DARPin promotes faster onset of botulinum neurotoxin A1 action. Nature Communications, 14, 8317. https://doi.org/10.1038/s41467-023-44102-4

Abstract

In this study, we characterize Designed Ankyrin Repeat Proteins (DARPins) as investigative tools to probe botulinum neurotoxin A1 (BoNT/A1) structure and function. We identify DARPin-F5 that completely blocks SNAP25 substrate cleavage by BoNT/A1 in vitro. X-ray crystallography reveals that DARPin-F5 inhibits BoNT/A1 activity by interacting with a substrate-binding region between the α- and β-exosite. This DARPin does not block substrate cleavage of BoNT/A3, indicating that DARPin-F5 is a subtype-specific inhibitor. BoNT/A1 Glu-171 pla