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Res Microbiol. 2018 Nov 20. pii: S0923-2508(18)30149-9. doi: 10.1016/j.resmic.2018.11.003. [Epub ahead of print]

Role of the Klebsiella pneumoniae TolC porin in antibiotic efflux.

Author information

1
Entasis Therapeutics, 35 Gatehouse Drive, Waltham, MA, USA. Electronic address: ram.iyer@entasistx.com.
2
Entasis Therapeutics, 35 Gatehouse Drive, Waltham, MA, USA. Electronic address: samir.moussa@entasistx.com.
3
Entasis Therapeutics, 35 Gatehouse Drive, Waltham, MA, USA. Electronic address: ruben.tommasi@entasistx.com.
4
Entasis Therapeutics, 35 Gatehouse Drive, Waltham, MA, USA. Electronic address: alita.miller@entasistx.com.

Abstract

The major Gram-negative gated efflux channel TolC has been extensively characterized in Escherichia coli but there is minimal information about Klebsiella pneumoniae TolC. Using an arabinose-inducible plasmid-based expression system, we show that the K. pneumoniae TolC complements the efflux defect in an E. coli K-12 ΔtolC strain, restoring wild-type levels of resistance towards most antibiotics suggesting that it can interact with the E. coli AcrB efflux pump. We characterize the efflux properties of K. pneumoniae TolC using an orthogonal whole cell-based assay and quantify the extrusion of environment-sensitive fluorescent probes and contrast the findings with the E. coli ortholog.

KEYWORDS:

E. coli; Efflux; Fluorescent dyes; K. pneumoniae; TolC

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