Studies on the inhibition of Escherichia coli Phospho-N-acetylmuramyl-pentapeptide Translocase
Studies on the inhibition of Escherichia coli Phospho-N-acetylmuramyl-pentapeptide Translocase
The problem of antibiotic resistance is an ever increasing threat to modern healthcare and there is an urgent need for the development of new antibacterial agents. Phospho-N-acetylmuramyl-pentapeptide translocase (translocase I) catalyses the first membrane step in the biosynthesis of the bacterial cell wall polymer peptidoglycan, and is a potential enzyme target for such novel agents. The mraY gene encoding for translocase I in E. coli has been cloned, overexpressed and the integral membrane protein solubilised with detergent.
Mureidomycin A (MRD A) is a peptidyl nucleoside antibiotic which has been characterised as a slow-binding inhibitor of translocase II (IC50 <0.1 μM). Selective modification of the primary amino and phenolic hydroxyl groups of MRD A has furnished derivatives which have been demonstrated to retain inhibitory activity against translocase I in a radiochemical assay (IC50 7.0-132 μM).
A uridine-based analogue of the unusual enamide moiety present in MRD A has been synthesised via aza-Wittig methodology. Experiments have demonstrated that this compound exhibits unexpected chemical stability, consistent with the observed inertness of the natural product. These results suggest that the enamide moiety is not involved in any active site chemistry, and leads to the hypothesis that the EI to EI* transition of slow-binding inhibition may be a conformational change.
Several potential translocase I inhibitors have been synthesised via the selective 5'-O-modification of uridine. Reversible enzyme inhibition by 5'-O-(ω-aminoacyl)uridines was shown to be dependent on the acyl chain length (3-aminopropanoyl-IC50 0.26 mM, 7-aminoheptanoyl- IC50 1.5 mM). There was also evidence for irreversible inhibition of translocase I by 5'-O-[3-{ethyl}phosphonate)propyl] uridine (IC50 3.7 mM). These compounds provide the basis for the rational design of more potent enzyme inhibitors.
University of Southampton
1998
Gentle, Caragh Ann
(1998)
Studies on the inhibition of Escherichia coli Phospho-N-acetylmuramyl-pentapeptide Translocase.
University of Southampton, Doctoral Thesis.
Record type:
Thesis
(Doctoral)
Abstract
The problem of antibiotic resistance is an ever increasing threat to modern healthcare and there is an urgent need for the development of new antibacterial agents. Phospho-N-acetylmuramyl-pentapeptide translocase (translocase I) catalyses the first membrane step in the biosynthesis of the bacterial cell wall polymer peptidoglycan, and is a potential enzyme target for such novel agents. The mraY gene encoding for translocase I in E. coli has been cloned, overexpressed and the integral membrane protein solubilised with detergent.
Mureidomycin A (MRD A) is a peptidyl nucleoside antibiotic which has been characterised as a slow-binding inhibitor of translocase II (IC50 <0.1 μM). Selective modification of the primary amino and phenolic hydroxyl groups of MRD A has furnished derivatives which have been demonstrated to retain inhibitory activity against translocase I in a radiochemical assay (IC50 7.0-132 μM).
A uridine-based analogue of the unusual enamide moiety present in MRD A has been synthesised via aza-Wittig methodology. Experiments have demonstrated that this compound exhibits unexpected chemical stability, consistent with the observed inertness of the natural product. These results suggest that the enamide moiety is not involved in any active site chemistry, and leads to the hypothesis that the EI to EI* transition of slow-binding inhibition may be a conformational change.
Several potential translocase I inhibitors have been synthesised via the selective 5'-O-modification of uridine. Reversible enzyme inhibition by 5'-O-(ω-aminoacyl)uridines was shown to be dependent on the acyl chain length (3-aminopropanoyl-IC50 0.26 mM, 7-aminoheptanoyl- IC50 1.5 mM). There was also evidence for irreversible inhibition of translocase I by 5'-O-[3-{ethyl}phosphonate)propyl] uridine (IC50 3.7 mM). These compounds provide the basis for the rational design of more potent enzyme inhibitors.
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Published date: 1998
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Local EPrints ID: 463534
URI: http://eprints.soton.ac.uk/id/eprint/463534
PURE UUID: 25a5a96b-e170-42fb-8e2b-a724b166dbbe
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Date deposited: 04 Jul 2022 20:53
Last modified: 04 Jul 2022 20:53
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Author:
Caragh Ann Gentle
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