CERTainly good to know!

Bioluminescence Resonance Energy Transfer (BRET) is a technique to observe and measure the interactions between proteins within living cells and a valuable tool for biomedical research. The Organic and Bioorganic Chemistry Group (Christoph Arenz) established a novel BRET system for identifying inhibitors of the lipid transfer protein CERT. By fusing Nanoluciferase to CERT and utilizing Nile red-labeled ceramide, they created a sensitive assay to monitor CERT activity in living cells. Using this innovative approach they were able to screen a combinatorial library of HPA-12 derivatives and identify six compounds with improved inhibitory activity compared to the parent compound. If you want to find out more about this valuable tool, check out their Angewandte Chemie International Edition Article!

Abstract

A BRET system is described, in which Nanoluciferase was fused to the lipid transfer protein CERT for efficient energy transfer to a Nile red-labeled ceramide, which is either directly bound to CERT or transported to the adjacent Golgi membrane. Bulk formation of sphingomyelin, a major plasma membrane component in mammals, is dependent on CERT-mediated transfer of its predecessor ceramide. CERT is considered a promising drug target but no direct cell-based methods exist to efficiently identify inhibitors. The utility of the method was demonstrated by a library of 140 derivatives of the CERT inhibitor HPA-12. These were obtained in a combinatorial synthesis using solid-phase transacylation. Screening of the library led to six compounds that were picked and confirmed to be superior to HPA-12 in a subsequent dose-response study and also in an orthogonal lipidomics analysis.