Publications

• Pyruvate-uridine diphospho-N-acetylglucosamine transferase. Purification to homogeneity and feedback inhibition.

  Zemell R.I., Anwar R.A.

  Phosphoenolpyruvate:uridine-5'-diphospho-N-acetyl-2-amino-2-deoxyglucose-3-enolpyruvyltranferase catalyzes the transfer of enolpyruvate from phosphoenolpyruvate to uridine diphospho-N-acetylglucosamine with the liberation of inorganic orthophosphate. It was purified to homogeneity from Enterobacte ... >> More

  Phosphoenolpyruvate:uridine-5'-diphospho-N-acetyl-2-amino-2-deoxyglucose-3-enolpyruvyltranferase catalyzes the transfer of enolpyruvate from phosphoenolpyruvate to uridine diphospho-N-acetylglucosamine with the liberation of inorganic orthophosphate. It was purified to homogeneity from Enterobacter cloacae with the use of UDP-N-acetylmuramyl-L-Ala-D-Glu-meso-Dap, a feedback inihibitor, as a ligand covalenty bound to Sepharose 4B. The evidence suggests that the enzyme is a single polypeptide with a molecular weight of 41,000. The enzyme catalyzes the first committed step in the biosynthesis of bacterial cell wall peptidoglycan. The cytoplasmic end product of this pathway is UDP-N-acetylmuramyl-L-Ala-D-Glu-meso-Dap-D-Ala-D-Ala (see article). UDP-MurNAc-pentapeptide and its precursor, UDP-MurNAc-tripeptide, were found to be effective inhibitiors of the enzyme. The kinetic data suggest a binding site for these inhibitors distinct from the active site. This is consistent with the proposed role for UDP-MurNAc-tripeptide and pentapeptide as negative modulators of the enzyme. << Less

  J Biol Chem 250:3185-3192(1975) [PubMed] [EuropePMC]

• Recent advances in the formation of the bacterial peptidoglycan monomer unit.

  van Heijenoort J.

  Nat Prod Rep 18:503-519(2001) [PubMed] [EuropePMC]

  This publication is cited by 11 other entries.

• Biosynthesis of uridine diphospho-N-acetylmuramic acid. II. Purification and properties of pyruvate-uridine diphospho-N-acetylglucosamine transferase and characterization of uridine diphospho-N-acetylenopyruvylglucosamine.

  Gunetileke K.G., Anwar R.A.

  J Biol Chem 243:5770-5778(1968) [PubMed] [EuropePMC]

• Cloning and sequencing of Escherichia coli murZ and purification of its product, a UDP-N-acetylglucosamine enolpyruvyl transferase.

  Marquardt J.L., Siegele D.A., Kolter R., Walsh C.T.

  The Escherichia coli gene murZ, encoding the enzyme UDP-N-acetylglucosamine enolpyruvyl transferase, has been cloned and sequenced. Identified by screening an E. coli genomic library for clones that conferred phosphomycin resistance, murZ encoded a 419-amino-acid polypeptide and was mapped to 69.3 ... >> More

  The Escherichia coli gene murZ, encoding the enzyme UDP-N-acetylglucosamine enolpyruvyl transferase, has been cloned and sequenced. Identified by screening an E. coli genomic library for clones that conferred phosphomycin resistance, murZ encoded a 419-amino-acid polypeptide and was mapped to 69.3 min on the E. coli chromosome. MurZ protein was purified to near homogeneity and found to have the expected UDP-N-acetylglucosamine enolpyruvyl transferase activity. Sequence analysis of the predicted product revealed 44% identity to OrfR from Bacillus subtilis (K. Trach, J.W. Chapman, P. Piggot, D. LeCoq, and J.A. Hoch, J. Bacteriol. 170:4194-4208, 1988), suggesting that orfR may also encode a UDP-N-acetylglucosamine enolpyruvyl transferase enzyme. MurZ is also homologous to the aromatic amino acid biosynthetic enzyme enolpyruvyl shikimate phosphate synthase, the other enzyme known to catalyze an enolpyruvyl transfer. << Less

  J. Bacteriol. 174:5748-5752(1992) [PubMed] [EuropePMC]

• The fungal product terreic acid is a covalent inhibitor of the bacterial cell wall biosynthetic enzyme UDP-N-acetylglucosamine 1-carboxyvinyltransferase (MurA).

  Han H., Yang Y., Olesen S.H., Becker A., Betzi S., Schoenbrunn E.

  Terreic acid is a metabolite with antibiotic properties produced by the fungus Aspergillus terreus. We found that terreic acid is a covalent inhibitor of the bacterial cell wall biosynthetic enzyme MurA from Enterobacter cloacae and Escherichia coli in vitro. The crystal structure of the MurA dead ... >> More

  Terreic acid is a metabolite with antibiotic properties produced by the fungus Aspergillus terreus. We found that terreic acid is a covalent inhibitor of the bacterial cell wall biosynthetic enzyme MurA from Enterobacter cloacae and Escherichia coli in vitro. The crystal structure of the MurA dead-end complex with terreic acid revealed that the quinine ring is covalently attached to the thiol group of Cys115, the molecular target of the antibiotic fosfomycin. Kinetic characterization established that the inactivation requires the presence of substrate UNAG (UDP-N-acetylglucosamine), proceeding with an inactivation rate constant k(inact) of 130 M(-1) s(-1). Although the mechanisms of inactivation are similar, fosfomycin is approximately 50 times more potent than terreic acid, and the structural consequences of covalent modification by these two inhibitors are fundamentally different. The MurA-fosfomycin complex exists in the closed enzyme conformation, with the Cys115-fosfomycin adduct buried in the active site. In contrast, the dead-end complex with terreic acid is open, is free of UNAG, and has the Cys115-terreic acid adduct solvent-exposed. It appears that terreic acid reacts with Cys115 in the closed, binary state of the enzyme, but that the resulting Cys115-terreic acid adduct imposes steric clashes in the active site. As a consequence, the loop containing Cys115 rearranges, the enzyme opens, and UNAG is released. The differential kinetic and structural characteristics of MurA inactivation by terreic acid and fosfomycin reflect the importance of noncovalent binding potential, even for covalent inhibitors, in ensuring inactivation efficiency and specificity. << Less

  Biochemistry 49:4276-4282(2010) [PubMed] [EuropePMC]