Biostructural analysis of the metal-sensor domain of CnrX from Cupriavidus metallidurans CH34

Antonie Van Leeuwenhoek. 2009 Aug;96(2):141-8. doi: 10.1007/s10482-008-9283-6. Epub 2008 Sep 30.

Abstract

In Cupriavidus metallidurans CH34, the proteins CnrX, CnrY, and CnrH regulate the expression of the cnrCBA operon that codes for a cation-efflux pump involved in cobalt and nickel resistance. The periplasmic part of CnrX can be defined as the metal sensor in the signal transduction complex composed of the membrane-bound anti-sigma factor CnrY and the extra-cytoplasmic function sigma factor CnrH. A soluble form of CnrX was overproduced and purified. This protein behaves as a dimer in solution as judged from gel filtration, sedimentation velocity experiments, and NMR. Native crystals diffracting to 2.3 A using synchrotron radiation were obtained using the hanging-drop vapor-diffusion method. They belong to the primitive monoclinic space group P2(1), with unit cell parameters a = 31.87, b = 74.80, c = 93.67 A, beta = 90.107 degrees. NMR data and secondary structure prediction suggest that this protein is essentially formed by helices.

MeSH terms

  • Amino Acid Sequence
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Cloning, Molecular
  • Crystallization
  • Cupriavidus / drug effects*
  • Cupriavidus / genetics
  • Cupriavidus / metabolism*
  • Dimerization
  • Drug Resistance, Bacterial*
  • Gene Expression Regulation, Bacterial
  • Magnetic Resonance Spectroscopy
  • Metals, Heavy / pharmacology*
  • Molecular Sequence Data
  • Periplasm / metabolism
  • Sequence Analysis, DNA
  • Signal Transduction*
  • Structure-Activity Relationship
  • X-Ray Diffraction

Substances

  • Bacterial Proteins
  • Metals, Heavy