4DN0

PelD 156-455 from Pseudomonas aeruginosa PA14 in complex with c-di-GMP

Summary for 4DN0

• Entry DOI: 10.2210/pdb4dn0/pdb
• Related: 4DMZ
• Descriptor: Putative uncharacterized protein pelD, 9,9'-[(2R,3R,3aS,5S,7aR,9R,10R,10aS,12S,14aR)-3,5,10,12-tetrahydroxy-5,12-dioxidooctahydro-2H,7H-difuro[3,2-d:3',2'-j][1,3,7,9,2,8]tetraoxadiphosphacyclododecine-2,9-diyl]bis(2-amino-1,9-dihydro-6H-purin-6-one), SULFATE ION, ... (5 entities in total)
• Functional Keywords: gaf domain, ggdef domain, c-di-gmp receptor, bacterial inner membrane, nucleotide-binding protein
• Biological source: Pseudomonas aeruginosa
• Total number of polymer chains: 1
• Total formula weight: 37582.86

Authors

Whitney, J.C.,Colvin, K.M.,Marmont, L.S.,Robinson, H.,Parsek, M.R.,Howell, P.L. (deposition date: 2012-02-08, release date: 2012-05-23, Last modification date: 2024-02-28)

Primary citation

Whitney, J.C.,Colvin, K.M.,Marmont, L.S.,Robinson, H.,Parsek, M.R.,Howell, P.L.
Structure of the Cytoplasmic Region of PelD, a Degenerate Diguanylate Cyclase Receptor That Regulates Exopolysaccharide Production in Pseudomonas aeruginosa.
J.Biol.Chem., 287:23582-23593, 2012

PubMed Abstract: High cellular concentrations of bis-(3',5')-cyclic dimeric guanosine mono-phosphate (c-di-GMP) regulate a diverse range of phenotypes in bacteria including biofilm development. The opportunistic pathogen Pseudomonas aeruginosa produces the PEL polysaccharide to form a biofilm at the air-liquid interface of standing cultures. Among the proteins required for PEL polysaccharide production, PelD has been identified as a membrane-bound c-di-GMP-specific receptor. In this work, we present the x-ray crystal structure of a soluble cytoplasmic region of PelD in its apo and c-di-GMP complexed forms. The structure of PelD reveals an N-terminal GAF domain and a C-terminal degenerate GGDEF domain, the latter of which binds dimeric c-di-GMP at an RXXD motif that normally serves as an allosteric inhibition site for active diguanylate cyclases. Using isothermal titration calorimetry, we demonstrate that PelD binds c-di-GMP with low micromolar affinity and that mutation of residues involved in binding not only decreases the affinity of this interaction but also abrogates PEL-specific phenotypes in vivo. Bioinformatics analysis of the juxtamembrane region of PelD suggests that it contains an α-helical stalk region that connects the soluble region to the transmembrane domains and that similarly to other GAF domain containing proteins, this region likely forms a coiled-coil motif that mediates dimerization. PelD with Alg44 and BcsA of the alginate and cellulose secretion systems, respectively, collectively constitute a group of c-di-GMP receptors that appear to regulate exopolysaccharide assembly at the protein level through activation of their associated glycosyl transferases.

PubMed: 22605337
DOI: 10.1074/jbc.M112.375378

Experimental method

X-RAY DIFFRACTION (2.3 Å)