An Efficient Way to Screen Inhibitors of Energy-Coupling Factor (ECF) Transporters in a Bacterial Uptake Assay.
Spyridon Bousis, Steffen Winkler, Jörg Haupenthal, Francesco Fulco, Eleonora Diamanti, Anna K H Hirsch
Author Information
Spyridon Bousis: Helmholtz Centre for Infection Research (HZI), Helmholtz Institute for Pharmaceutical Research (HIPS), Campus Building E 8.1, D-66123 Saarbrücken, Germany.
Steffen Winkler: Helmholtz Centre for Infection Research (HZI), Helmholtz Institute for Pharmaceutical Research (HIPS), Campus Building E 8.1, D-66123 Saarbrücken, Germany.
Jörg Haupenthal: Helmholtz Centre for Infection Research (HZI), Helmholtz Institute for Pharmaceutical Research (HIPS), Campus Building E 8.1, D-66123 Saarbrücken, Germany.
Francesco Fulco: Helmholtz Centre for Infection Research (HZI), Helmholtz Institute for Pharmaceutical Research (HIPS), Campus Building E 8.1, D-66123 Saarbrücken, Germany.
Eleonora Diamanti: Helmholtz Centre for Infection Research (HZI), Helmholtz Institute for Pharmaceutical Research (HIPS), Campus Building E 8.1, D-66123 Saarbrücken, Germany. ORCID
Anna K H Hirsch: Helmholtz Centre for Infection Research (HZI), Helmholtz Institute for Pharmaceutical Research (HIPS), Campus Building E 8.1, D-66123 Saarbrücken, Germany. ORCID
Herein, we report a novel whole-cell screening assay using as a model microorganism to identify inhibitors of energy-coupling factor (ECF) transporters. This promising and underexplored target may have important pharmacological potential through modulation of vitamin homeostasis in bacteria and, importantly, it is absent in humans. The assay represents an alternative, cost-effective and fast solution to demonstrate the direct involvement of these membrane transporters in a native biological environment rather than using a low-throughput in vitro assay employing reconstituted proteins in a membrane bilayer system. Based on this new whole-cell screening approach, we demonstrated the optimization of a weak hit compound () into a small molecule () with improved in vitro and whole-cell activities. This study opens the possibility to quickly identify novel inhibitors of ECF transporters and optimize them based on structure-activity relationships.
