Selection of peptide inhibitors against the Pseudomonas aeruginosa MurD cell wall enzyme
| Autor: | Lydia Boudreault, Mélanie Beaumont, Catherine Paradis-Bleau, Timothy D. H. Bugg, François Sanschagrin, Roger C. Levesque, Adrian J. Lloyd |
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| Rok vydání: | 2006 |
| Předmět: |
Phage display
Protein Conformation Physiology Amino Acid Motifs Molecular Sequence Data Peptide Biopanning Biology Biochemistry Ligases Inhibitory Concentration 50 Cellular and Molecular Neuroscience Endocrinology Protein structure Peptide Library Amino Acid Sequence Peptide library Peptide sequence chemistry.chemical_classification DNA ligase Binding Sites Dose-Response Relationship Drug Sequence Homology Amino Acid Computational Biology Sequence Analysis DNA Amino acid Models Chemical chemistry Spectrophotometry Pseudomonas aeruginosa Peptides Protein Binding |
| Zdroj: | Peptides. 27:1693-1700 |
| ISSN: | 0196-9781 |
| DOI: | 10.1016/j.peptides.2006.01.017 |
| Popis: | The purified Pseudomonas aeruginosa cell wall biosynthesis MurD amide ligase enzyme was used to screen C-7-C and 12 mers peptides from phage display libraries using competitive biopanning approaches with the specific substrates D-glutamate and ATP. From the 60 phage-encoded peptides identified, DNA was sequenced, deduced amino acid sequences aligned and two peptides were synthesized from consensus sequences identified. The UDP-N-acetylmuramyl-L-alanine MurD substrate was synthesized, purified and used to develop a spectrophotometric assay. One peptide synthesized was found to specifically inhibit ATPase activity of MurD. The IC50 value was estimated at 4 microM for the C-7-C MurDp1 peptide. The loop conformation of MurDp1 was shown to be important for the inhibition of the UDP-N-acetylmuramyl-L-alanine:D-glutamate MurD ligase. The linear 12 mers MurD2 peptide has an IC50 value of 15 mM. A conserved amino acid motif was found between MurDp2 and the bacterial glyceraldehyde 3-phosphate dehydrogenase indicating that MurDp2 binds at a protein-protein interacting site. The approach proposed and results obtained suggest that efficient peptide inhibitors as well as protein-protein interaction domains can be identified by phage display. |
| Databáze: | OpenAIRE |
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