title: Crystal structure of a catalytically active GG(D/E)EF diguanylate cyclase domain from Marinobacter aquaeolei with bound c-di-GMP product.
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.
authors: Department of Biological Sciences, The Northeast Structural Genomics Consortium Columbia University, New York, NY, 10032, USA.
authors: Sergey M
authors: SM
authors: Vorobiev
authors: Helen
authors: H
authors: Neely
authors: Bomina
authors: B
authors: Yu
authors: Jayaraman
authors: J
authors: Seetharaman
authors: Rong
authors: R
authors: Xiao
authors: Thomas B
authors: TB
authors: Acton
authors: Gaetano T
authors: GT
authors: Montelione
authors: John F
authors: JF
authors: Hunt
mesh_terms: D000494:Allosteric Regulation; D000495:Allosteric Site; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry*; D017124:Conserved Sequence; D018360:Crystallography, X-Ray / Q000379:methods; D006152:Cyclic GMP / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000737:chemistry; D004789:Enzyme Activation; D029968:Escherichia coli Proteins / Q000737:chemistry*; D006160:Guanosine Triphosphate / Q000737:chemistry; D053520:Marinobacter / Q000737:chemistry* / Q000201:enzymology; D008969:Molecular Sequence Data; D019761:Phosphorus-Oxygen Lyases / Q000737:chemistry*; D011487:Protein Conformation; D025941:Protein Interaction Mapping; D017434:Protein Structure, Tertiary; D020539:Sequence Analysis, Protein
chemical_list: D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; C062025:bis(3',5')-cyclic diguanylic acid; D006160:Guanosine Triphosphate; D019761:Phosphorus-Oxygen Lyases; C060720:diguanylate cyclase; D006152:Cyclic GMP