Hunting enzymes in the membrane: A new strategy to inhibit rhomboid proteases

The researchers led by Kvido Stříšovský from IOCB Prague and Adam Lange from Leibniz Forschungsinstitut für Molekulare Pharmakologie in Berlin built a high-throughput screening platform that mimics the natural membrane setting and uses a native-like substrate. This physiologically relevant setup led to the discovery of several chemically diverse, non-peptidic inhibitors that selectively block the bacterial rhomboid GlpG protease, which is a key pathogenic factor in uropathogenic, biofilm-forming bacteria.

One of the active compounds showed particularly promising potency against both GlpG and the human mitochondrial rhomboid PARL, which regulates mitochondrial quality control and stress responses and has been implicated in the pathogenesis of Parkinson’s disease. Analogues of another hit compound revealed a new class of reversible covalent inhibitors that even function in living bacteria with permeabilized membranes. By combining docking studies, structural analysis of binding mode by solid-state NMR, and structure–activity analysis aided by computational modeling, the joint international team uncovered how these molecules bind and interact with critical catalytic features of the enzyme.

Beyond identifying new inhibitor scaffolds, the study provides a blueprint for targeting rhomboid proteases in disease contexts such as Parkinson’s disease, malaria, bacterial biofilms in the urinary tract and other infections, which is an important step toward future therapeutic applications.

• Bohg, C.; Dubanych, Y.; Kosteletos, S.; et al. Targeting the Membrane-Embedded Rhomboid Protease GlpG: A Multimodal Strategy for Inhibitor Discovery and Mechanistic Insight. Angew. Chem. Int. Ed. 2026, e14067. https://doi.org/10.1002/anie.202514067