Slow binding inhibition of phospho-N-acetylmuramyl-pentapeptide-translocase (Escherichia coli) by mureidomycin A.
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.
Brandish
P E
PE
Department of Chemistry, Southampton University, Highfield, United Kingdom.
Burnham
M K
MK
Lonsdale
J T
JT
Southgate
R
R
Inukai
M
M
Bugg
T D
TD
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
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