Crystal structure of an EAL domain in complex with reaction product 5'-pGpG

PLoS One. 2012;7(12):e52424. doi: 10.1371/journal.pone.0052424. Epub 2012 Dec 20.

Abstract

FimX is a large multidomain protein containing an EAL domain and involved in twitching motility in Pseudomonas aeruginosa. We present here two crystallographic structures of the EAL domain of FimX (residues 438-686): one of the apo form and the other of a complex with 5'-pGpG, the reaction product of the hydrolysis of c-di-GMP. In both crystal forms, the EAL domains form a dimer delimiting a large cavity encompassing the catalytic pockets. The ligand is trapped in this cavity by its sugar phosphate moiety. We confirmed by NMR that the guanine bases are not involved in the interaction in solution. We solved here the first structure of an EAL domain bound to the reaction product 5'-pGpG. Though isolated FimX EAL domain has a very low catalytic activity, which would not be significant compared to other catalytic EAL domains, the structure with the product of the reaction can provides some hints in the mechanism of hydrolysis of the c-di-GMP by EAL domains.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Apoproteins / chemistry
  • Apoproteins / metabolism
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / metabolism*
  • Catalytic Domain
  • Crystallography, X-Ray
  • Cyclic GMP / analogs & derivatives
  • Cyclic GMP / metabolism
  • Deoxyguanine Nucleotides / metabolism*
  • Hydrolysis
  • Ligands
  • Magnesium / metabolism
  • Models, Molecular
  • Molecular Sequence Data
  • Nitrophenols / metabolism
  • Phosphoric Diester Hydrolases / metabolism
  • Protein Binding
  • Protein Multimerization
  • Protein Structure, Tertiary
  • Pseudomonas aeruginosa / enzymology*
  • Sequence Alignment
  • Solutions

Substances

  • Apoproteins
  • Bacterial Proteins
  • Deoxyguanine Nucleotides
  • Ligands
  • Nitrophenols
  • Solutions
  • deoxydiguanosine diphosphosphate
  • bis(3',5')-cyclic diguanylic acid
  • bis(4-nitrophenyl)phosphate
  • Phosphoric Diester Hydrolases
  • Cyclic GMP
  • Magnesium

Associated data

  • PDB/4AFY
  • PDB/4AG0

Grants and funding

This work was funded by CNRS. Publication costs were supported by the Institut Universitaire de France. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.