title: Detection and function of an intramolecular disulfide bond in the pH-responsive CadC of Escherichia coli.
abstract: BACKGROUND
  In an acidic and lysine-rich environment Escherichia coli induces expression of the cadBA operon which encodes CadA, the lysine decarboxylase, and CadB, the lysine/cadaverine antiporter. cadBA expression is dependent on CadC, a membrane-integrated transcriptional activator which belongs to the ToxR-like protein family. Activation of CadC requires two stimuli, lysine and low pH. Whereas lysine is detected by an interplay between CadC and the lysine-specific transporter LysP, pH alterations are sensed by CadC directly. Crystal structural analyses revealed a close proximity between two periplasmic cysteines, Cys208 and Cys272.
  
  
  RESULTS
  Substitution of Cys208 and/or Cys272 by alanine resulted in CadC derivatives that were active in response to only one stimulus, either lysine or pH 5.8. Differential in vivo thiol trapping revealed a disulfide bond between these two residues at pH 7.6, but not at pH 5.8. When Cys208 and Cys272 were replaced by aspartate and lysine, respectively, virtually wild-type behavior was restored indicating that the disulfide bond could be mimicked by a salt bridge.
  
  
  CONCLUSIONS
  A disulfide bond was found in the periplasmic domain of CadC that supports an inactive state of CadC at pH 7.6. At pH 5.8 disulfide bond formation is prevented which transforms CadC into a semi-active state. These results provide new insights into the function of a pH sensor.
authors.affiliation: Munich Center for integrated Protein Science, Department of Biology I, Microbiology, Ludwig-Maximilians-Universität München, Grosshaderner Straße 2-4, 82152 Martinsried, Germany.
authors.forename: Larissa
authors.initials: L
authors.lastname: Tetsch
authors.forename: Christiane
authors.initials: C
authors.lastname: Koller
authors.forename: Alexandra
authors.initials: A
authors.lastname: Dönhöfer
authors.forename: Kirsten
authors.initials: K
authors.lastname: Jung
mesh_terms: D019943:Amino Acid Substitution / Q000235:genetics; D003545:Cysteine / Q000235:genetics; D004220:Disulfides / Q000737:chemistry* / Q000378:metabolism*; D004926:Escherichia coli / Q000737:chemistry* / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008239:Lysine / Q000378:metabolism; D008954:Models, Biological; D008956:Models, Chemical; D016297:Mutagenesis, Site-Directed; D050505:Mutant Proteins / Q000737:chemistry / Q000378:metabolism; D015534:Trans-Activators / Q000737:chemistry* / Q000378:metabolism*
chemical_list: C087479:CadC protein, E coli; D004220:Disulfides; D029968:Escherichia coli Proteins; D050505:Mutant Proteins; D015534:Trans-Activators; D008239:Lysine; D003545:Cysteine