Cloning, purification, crystallization and X-ray crystallographic analysis of the periplasmic sensing domain of Pseudomonas fluorescens chemotactic transducer of amino acids type A (CtaA)

Biosci Trends. 2016 Sep 5;10(4):320-4. doi: 10.5582/bst.2016.01059. Epub 2016 Jun 2.

Abstract

Chemotaxis towards nutrients plays a crucial role in root colonization by Pseudomonas fluorescens. The P. fluorescens chemotactic transducer of amino acids type A (CtaA) mediates movement towards amino acids present in root exudates. In this study, the periplasmic sensory domain of CtaA has been crystallized by the hanging-drop vapor diffusion method using ammonium sulfate as a precipitating agent. A complete data set was collected to 1.9 Å resolution using cryocooling conditions and synchrotron radiation. The crystals belong to space group I222 or I212121, with unit-cell parameters a = 67.2, b = 76.0, c = 113.3 Å. This is an important step towards elucidation of the structural basis of how CtaA recognizes its signal molecules and transduces the signal across the membrane.

MeSH terms

  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / isolation & purification
  • Chemotaxis / genetics*
  • Cloning, Molecular
  • Crystallization / methods
  • Crystallography, X-Ray
  • Cytochrome b Group / chemistry*
  • Cytochrome b Group / isolation & purification
  • Membrane Proteins / chemistry*
  • Membrane Proteins / isolation & purification
  • Plant Roots / microbiology
  • Protein Domains
  • Pseudomonas fluorescens / chemistry*
  • Signal Transduction

Substances

  • Bacterial Proteins
  • CtaA protein, bacteria
  • Cytochrome b Group
  • Membrane Proteins