| Autor: |
Carson DV; Department of Chemical and Biological Engineering, Princeton University, Princeton, New Jersey 08544, United States., Juarez RJ; Chemical and Physical Biology Program, Vanderbilt University, Nashville, Tennessee 37235, United States., Do T; Department of Chemical and Biological Engineering, Princeton University, Princeton, New Jersey 08544, United States., Yang ZJ; Department of Chemistry, Vanderbilt University, Nashville, Tennessee 37235, United States.; Center for Structural Biology, Vanderbilt University, Nashville, Tennessee 37235, United States.; Vanderbilt Institute of Chemical Biology, Vanderbilt University, Nashville, Tennessee 37235, United States.; Data Science Institute, Vanderbilt University, Nashville, Tennessee 37235, United States.; Department of Chemical and Biomolecular Engineering, Vanderbilt University, Nashville, Tennessee 37235, United States., Link AJ; Department of Chemical and Biological Engineering, Princeton University, Princeton, New Jersey 08544, United States.; Department of Chemistry, Princeton University, Princeton, New Jersey 08544, United States.; Department of Molecular Biology, Princeton University, Princeton, New Jersey 08544, United States. |
| Abstrakt: |
The development of new antimicrobial agents effective against Gram-negative bacteria remains a major challenge in drug discovery. The lasso peptide cloacaenodin has potent antimicrobial activity against multiple strains in the Enterobacter genus, one of the ESKAPE pathogens. Here, we show that cloacaenodin uses a previously uncharacterized TonB-dependent transporter, which we name CloU, to cross the outer membrane (OM) of susceptible bacteria. Inner membrane transport is mediated by the protein SbmA. CloU is distinct from the known OM transporters (FhuA and PupB) utilized by other antimicrobial lasso peptides and thus offers important insight into the spectrum of activity of cloacaenodin. Using knowledge of the transport pathway to predict other cloacaenodin-susceptible strains, we demonstrate the activity of cloacaenodin against clinical isolates of Enterobacter and of a Kluyvera strain. Further, we use molecular dynamics simulations and mutagenesis of CloU to explain the variation in cloacaenodin susceptibility observed across different strains of Enterobacter . This work expands the currently limited understanding of lasso peptide uptake and advances the potential of cloacaenodin as an antibiotic. |
