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Development of a dual chemical probe for the USP16 and HDAC6 zinc-finger ubiquitin-binding domain

Created on 24 Oct 2025

Authors

Santhakumar, V., Silva, M., Mann, M. K., Mirabi, B., Szewczyk, M., Loup, J., Dupeux, A., Chandrasekaran, R., Bajohr, J., Arrowsmith, C., Schapira, M., Lautens, M., Barsyte-Lovejoy, D., Harding, R. J.

Abstract

Ubiquitin-specific peptidase 16 (USP16) is a deubiquitinase that specifically cleaves ubiquitin from histone H2A, and modulates gene expression, cell cycle regulation, and various other cellular processes. The USP16 zinc-finger ubiquitin-binding domain (UBD) binds the free C-terminal end of both ubiquitin and ISG15, two major signaling proteins that mediate many biological pathways. Because the precise function of USP16-UBD and its interactions remains unclear, a small molecule antagonist targeting the USP16-UBD could enable cellular studies to elucidate its biological role. Here we report SGC-UBD1031 (15), a chemical probe targeting USP16-UBD with similar in vitro binding profiles to HDAC6-UBD and selectivity over nine other UBDs. In cellular assays, 15 disrupts the interaction between the C-terminus of ISG15 and USP1-UBD, as well as the interaction between ISG15 and HDAC6 UBD, at a concentration of 1 M. The corresponding enantiomer SGC-UBD1031N (16), does not interfere with these interactions, even at concentrations as high as 30 M, and thus serves as a negative control.

Preprint server: bioRxiv
The authors list and abstract were imported from bioRxiv on 24 Oct 2025.

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