Application of Synthetic Peptide Arrays To Uncover Cyclic Di-GMP Binding Motifs
Cast your vote
You can rate an item by clicking the amount of stars they wish to award to this item.
When enough users have cast their vote on this item, the average rating will also be shown.
Your vote was cast
Thank you for your feedback
Thank you for your feedback
Herberg, Friedrich W.
MetadataShow full item record
AbstractABSTRACT High levels of the universal bacterial second messenger cyclic di-GMP (c-di-GMP) promote the establishment of surface-attached growth in many bacteria. Not only can c-di-GMP bind to nucleic acids and directly control gene expression, but it also binds to a diverse array of proteins of specialized functions and orchestrates their activity. Since its development in the early 1990s, the synthetic peptide array technique has become a powerful tool for high-throughput approaches and was successfully applied to investigate the binding specificity of protein-ligand interactions. In this study, we used peptide arrays to uncover the c-di-GMP binding site of a Pseudomonas aeruginosa protein (PA3740) that was isolated in a chemical proteomics approach. PA3740 was shown to bind c-di-GMP with a high affinity, and peptide arrays uncovered LKKALKKQTNLR to be a putative c-di-GMP binding motif. Most interestingly, different from the previously identified c-di-GMP binding motif of the PilZ domain (RXXXR) or the I site of diguanylate cyclases (RXXD), two leucine residues and a glutamine residue and not the charged amino acids provided the key residues of the binding sequence. Those three amino acids are highly conserved across PA3740 homologs, and their singular exchange to alanine reduced c-di-GMP binding within the full-length protein. IMPORTANCE In many bacterial pathogens the universal bacterial second messenger c-di-GMP governs the switch from the planktonic, motile mode of growth to the sessile, biofilm mode of growth. Bacteria adapt their intracellular c-di-GMP levels to a variety of environmental challenges. Several classes of c-di-GMP binding proteins have been structurally characterized, and diverse c-di-GMP binding domains have been identified. Nevertheless, for several c-di-GMP receptors, the binding motif remains to be determined. Here we show that the use of a synthetic peptide array allowed the identification of a c-di-GMP binding motif of a putative c-di-GMP receptor protein in the opportunistic pathogen P. aeruginosa . The application of synthetic peptide arrays will facilitate the search for additional c-di-GMP receptor proteins and aid in the characterization of c-di-GMP binding motifs.
CitationApplication of Synthetic Peptide Arrays To Uncover Cyclic Di-GMP Binding Motifs 2016, 198 (1):138 Journal of Bacteriology
AffiliationHelmholtz Centre for infection research, Inhoffenstr. 7, D-38124 Braunschweig, Germany.
JournalJournal of Bacteriology
The following license files are associated with this item:
- Application of Synthetic Peptide Arrays To Uncover Cyclic Di-GMP Binding Motifs.
- Authors: Düvel J, Bense S, Möller S, Bertinetti D, Schwede F, Morr M, Eckweiler D, Genieser HG, Jänsch L, Herberg FW, Frank R, Häussler S
- Issue date: 2016 Jan 1
- The second messenger bis-(3'-5')-cyclic-GMP and its PilZ domain-containing receptor Alg44 are required for alginate biosynthesis in Pseudomonas aeruginosa.
- Authors: Merighi M, Lee VT, Hyodo M, Hayakawa Y, Lory S
- Issue date: 2007 Aug
- Structural Conservation and Diversity of PilZ-Related Domains.
- Authors: Galperin MY, Chou SH
- Issue date: 2020 Jan 29
- Structure of the cytoplasmic region of PelD, a degenerate diguanylate cyclase receptor that regulates exopolysaccharide production in Pseudomonas aeruginosa.
- Authors: Whitney JC, Colvin KM, Marmont LS, Robinson H, Parsek MR, Howell PL
- Issue date: 2012 Jul 6
- PilZ Domain Protein FlgZ Mediates Cyclic Di-GMP-Dependent Swarming Motility Control in Pseudomonas aeruginosa.
- Authors: Baker AE, Diepold A, Kuchma SL, Scott JE, Ha DG, Orazi G, Armitage JP, O'Toole GA
- Issue date: 2016 Jul 1