Differential role of calmodulin and calcium ions in the stabilization of the catalytic domain of adenyl cyclase CyaA from Bordetella pertussis

Proteins. 2012 Apr;80(4):1028-40. doi: 10.1002/prot.24005. Epub 2012 Jan 9.

Abstract

The catalytic adenyl cyclase (AC) domain of the protein CyaA from Bordetella pertussis is activated by interaction with the C terminal lobe of calmodulin (C-CaM). The AC/C-CaM complex displays an elongated shape, but hydrodynamics measurements on the isolated AC domain allowed to characterize the shape of the protein as spherical. Here, we study by molecular dynamics simulations the complexes between AC and the apo and Ca(2+)-loaded C-CaM, as well as the isolated AC, to characterize the features of AC conformational variability and of AC/C-CaM interaction. The removal of calcium ions from C-CaM increases the AC flexibility, but the removal of C-CaM induces a dramatic drift of the AC conformation. Isolated AC conformations show a general tendency to become less elongated, as the two protein extremities (regions SA and CB) tend to get closer. An analysis of the energetic influences between the C-CaM and the AC regions shows a simple influence scheme, in agreement with the high affinity of AC to CaM. In this scheme, a single influence is observed from C-CaM to the region CA of the AC domain. This influence is correlated to the presence of hydrogen bonds involving residues from C-CaM, and from regions CA, C-terminal tail, and catalytic loop of AC. This study reveals a C-CaM/AC interaction picture where C-CaM stabilizes AC by a steric hindrance on the conformational drift of SA, whereas the Ca(2+) ions allow further stabilization by the establishment of a hydrogen bond network extending from C-CaM to the AC catalytic loop.

MeSH terms

  • Adenylate Cyclase Toxin / chemistry*
  • Bacterial Proteins / chemistry
  • Bordetella pertussis / chemistry
  • Bordetella pertussis / enzymology*
  • Calcium / chemistry*
  • Calmodulin / chemistry*
  • Catalytic Domain*
  • Crystallography, X-Ray
  • Enzyme Activation
  • Enzyme Stability
  • Hydrogen Bonding
  • Molecular Dynamics Simulation
  • Principal Component Analysis
  • Protein Interaction Mapping

Substances

  • Adenylate Cyclase Toxin
  • Bacterial Proteins
  • Calmodulin
  • Calcium