Lighting up the linear Ubiquitin Code: A Bioluminescent Sensor for M1-Specific Deubiquitinase Activity and Drug Discovery

Abstract

A luciferase-based reporter, N-Rluc-Ub-Ub-C-Rluc was developed to selectively monitor M1-deubiquitinase activity via catalytic signal amplification. This high-throughput assay identified Baicalein and Lumacaftor as novel OTULIN inhibitors from an FDA-approved compound library, validated by STD-NMR and cellular assays. The platform enables drug discovery targeting linear ubiquitin signaling without specialized synthesis.