Crystal structure of a catalytically active GG(D/E)EF diguanylate cyclase domain from Marinobacter aquaeolei with bound c-di-GMP product

J Struct Funct Genomics. 2012 Sep;13(3):177-83. doi: 10.1007/s10969-012-9136-4. Epub 2012 Jul 29.

Abstract

Recent studies of signal transduction in bacteria have revealed a unique second messenger, bis-(3'-5')-cyclic dimeric GMP (c-di-GMP), which regulates transitions between motile states and sessile states, such as biofilms. C-di-GMP is synthesized from two GTP molecules by diguanylate cyclases (DGC). The catalytic activity of DGCs depends on a conserved GG(D/E)EF domain, usually part of a larger multi-domain protein organization. The domains other than the GG(D/E)EF domain often control DGC activation. This paper presents the 1.83 Å crystal structure of an isolated catalytically competent GG(D/E)EF domain from the A1U3W3_MARAV protein from Marinobacter aquaeolei. Co-crystallization with GTP resulted in enzymatic synthesis of c-di-GMP. Comparison with previously solved DGC structures shows a similar orientation of c-di-GMP bound to an allosteric regulatory site mediating feedback inhibition of the enzyme. Biosynthesis of c-di-GMP in the crystallization reaction establishes that the enzymatic activity of this DGC domain does not require interaction with regulatory domains.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Allosteric Regulation
  • Allosteric Site
  • Amino Acid Sequence
  • Bacterial Proteins / chemistry*
  • Conserved Sequence
  • Crystallography, X-Ray / methods
  • Cyclic GMP / analogs & derivatives*
  • Cyclic GMP / biosynthesis
  • Cyclic GMP / chemistry
  • Enzyme Activation
  • Escherichia coli Proteins / chemistry*
  • Guanosine Triphosphate / chemistry
  • Marinobacter / chemistry*
  • Marinobacter / enzymology
  • Molecular Sequence Data
  • Phosphorus-Oxygen Lyases / chemistry*
  • Protein Conformation
  • Protein Interaction Mapping
  • Protein Structure, Tertiary
  • Sequence Analysis, Protein

Substances

  • Bacterial Proteins
  • Escherichia coli Proteins
  • bis(3',5')-cyclic diguanylic acid
  • Guanosine Triphosphate
  • Phosphorus-Oxygen Lyases
  • diguanylate cyclase
  • Cyclic GMP