Biophysical Society Conference | Tahoe 2022

Molecular Biophysics of Membranes

Thursday Speaker Abstracts

COLICIN E1 OPENS ITS HINGE TO PLUG TOLC Joanna S.G. Slusky 1,2 ; Jimmy Budiardjo 1 ; Jacqueline J Stevens 2 ; Anna Calkins 3 ; Ayotunde P Ikujuni 2 ; Virangika K Wimalasena 2 ; Emre Firlar 4 ; David A Case 5 ; Julie S Biteen 3 ; Jason T Kaelber 4 ; 1 University of Kansas, Center for Computational Biology, Lawrence, KS, USA 2 University of Kansas, Department of Molecular Biosciences, Lawrence, KS, USA 3 University of Michigan, Department of Chemistry, Ann Arbor , MI, USA 4 Rutgers University, Rutgers CryoEM & Nanoimaging Facility and Institute for Quantitative Biomedicine, Piscataway, NJ, USA 5 Rutgers University, Department of Chemistry and Chemical Biology, Piscataway, NJ, USA The double membrane architecture of Gram-negative bacteria forms a barrier that is impermeable to most extracellular threats. Bacteriocin proteins evolved to exploit the accessible, surface-exposed proteins embedded in the outer membrane to deliver cytotoxic cargo. Colicin E1 is a bacteriocin produced by, and lethal to, Escherichia coli that hijacks the outer membrane proteins TolC and BtuB to enter the cell. Here we capture the colicin E1 translocation domain inside its membrane receptor, TolC, by high-resolution cryoEM to obtain the first reported structure of a bacteriocin bound to TolC. Colicin E1 binds stably to TolC as an open hinge through the TolC pore—an architectural rearrangement from colicin E1’s unbound conformation. This binding is stable in live E. coli cells as indicated by single-molecule fluorescence microscopy. Finally, colicin E1 fragments binding to TolC plug the channel, inhibiting its native efflux function as an antibiotic efflux pump and heightening susceptibility to three antibiotic classes. In addition to demonstrating that these protein fragments are useful starting points for developing novel antibiotic potentiators, this method could be expanded to other colicins to inhibit other outer membrane protein functions.

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