title: Slow binding inhibition of phospho-N-acetylmuramyl-pentapeptide-translocase (Escherichia coli) by mureidomycin A.
abstract: Enzymes of the membrane cycle of reactions in bacterial peptidoglycan biosynthesis remain as unexploited potential targets for antibacterial agents. The first of these enzymes, phospho-N-acetylmuramyl-pentapeptide-translocase (EC 2.7.8.13), has been overexpresed in Escherichia coli and solubilized from particulate fractions. The work of W.A. Weppner and F.C. Neuhaus ((1977) J. Biol. Chem. 252, 2296-303) has been extended to establish a usable routine fluorescence-based continuous assay for solubilized preparations. This assay has been used in the characterization of the natural product, mureidomycin A as a potent slow binding inhibitor of the enzyme with Ki and Ki* of 36 nM and 2 nM, respectively.
authors.affiliation: Department of Chemistry, Southampton University, Highfield, United Kingdom.
authors.forename: P E
authors.initials: PE
authors.lastname: Brandish
authors.forename: M K
authors.initials: MK
authors.lastname: Burnham
authors.forename: J T
authors.initials: JT
authors.lastname: Lonsdale
authors.forename: R
authors.initials: R
authors.lastname: Southgate
authors.forename: M
authors.initials: M
authors.lastname: Inukai
authors.forename: T D
authors.initials: TD
authors.lastname: Bugg
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004926:Escherichia coli / Q000201:enzymology*; D005456:Fluorescent Dyes; D005822:Genetic Vectors; D007536:Isomerism; D007700:Kinetics; D008962:Models, Theoretical; D009705:Nucleosides / Q000494:pharmacology; D010957:Plasmids; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000302:isolation & purification; D013050:Spectrometry, Fluorescence; D013379:Substrate Specificity; D017855:Transferases (Other Substituted Phosphate Groups) / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000302:isolation & purification
chemical_list: D000900:Anti-Bacterial Agents; D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; D009705:Nucleosides; D011994:Recombinant Proteins; C059803:mureidomycin A; D017855:Transferases (Other Substituted Phosphate Groups); C100130:phospho-N-acetylmuramoyl pentapeptide transferase