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Role of the Klebsiella pneumoniae TolC porin in antibiotic efflux
Affiliation:1. Biozentrum, University Hospital Basel, Switzerland;2. Clinical Microbiology, University Hospital Basel, Switzerland;3. Applied Microbiology Research, Department of Biomedicine, University of Basel, CH-4056 Basel, Switzerland;4. BioVersys AG, Hochbergerstrasse 60C, Technology Park, 4057 Basel, Switzerland
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:TolC  Efflux  Fluorescent dyes  EV"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  kwrd0040"  },"  $$"  :[{"  #name"  :"  text"  ,"  _"  :"  Empty Vector  MIC"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  kwrd0050"  },"  $$"  :[{"  #name"  :"  text"  ,"  _"  :"  Minimum Inhibitory Concentration  NR"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  kwrd0060"  },"  $$"  :[{"  #name"  :"  text"  ,"  _"  :"  Nile Red  EtBr"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  kwrd0070"  },"  $$"  :[{"  #name"  :"  text"  ,"  _"  :"  Ethidium Bromide  Acr"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  kwrd0080"  },"  $$"  :[{"  #name"  :"  text"  ,"  _"  :"  Acriflavine  Ben"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  kwrd0090"  },"  $$"  :[{"  #name"  :"  text"  ,"  _"  :"  bis-benzamide  Ec_TolC"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  kwrd0100"  },"  $$"  :[{"  #name"  :"  text"  ,"  $$"  :[{"  #name"  :"  italic"  ,"  _"  :"  E. coli"  },{"  #name"  :"  __text__"  ,"  _"  :"   TolC  Kp_TolC"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  kwrd0110"  },"  $$"  :[{"  #name"  :"  text"  ,"  $$"  :[{"  #name"  :"  italic"  ,"  _"  :"  K. pneumoniae"  },{"  #name"  :"  __text__"  ,"  _"  :"   TolC
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