A Phosphinate Inhibitor of themeso-Diaminopimelic Acid-Adding Enzyme (MurE) of Peptidoglycan Biosynthesis

Binqi Zeng; Kenny K. Wong; David L. Pompliano; Sreelatha Reddy; Martin E. Tanner

doi:10.1021/jo981895p

Expression of the Staphylococcus aureusUDP-N-Acetylmuramoyl- l-Alanyl-d-Glutamate:l-Lysine Ligase in Escherichia coli and Effects on Peptidoglycan Biosynthesis and Cell Growth

Journal of Bacteriology ◽

10.1128/jb.181.19.5909-5914.1999 ◽

1999 ◽

Vol 181 (19) ◽

pp. 5909-5914 ◽

Cited By ~ 45

Author(s):

Dominique Mengin-Lecreulx ◽

Tim Falla ◽

Didier Blanot ◽

Jean van Heijenoort ◽

David J. Adams ◽

...

Keyword(s):

Escherichia Coli ◽

Morphological Changes ◽

Diaminopimelic Acid ◽

Side Chain ◽

E Coli ◽

Peptidoglycan Biosynthesis ◽

Additional Information ◽

The Third ◽

Composition And Structure ◽

Ca 50

ABSTRACT The monomer units in the Escherichia coli andStaphylococcus aureus cell wall peptidoglycans differ in the nature of the third amino acid in thel-alanyl-γ-d-glutamyl-X-d-alanyl-d-alanine side chain, where X is meso-diaminopimelic acid orl-lysine, respectively. The murE gene fromS. aureus encoding the UDP-N-acetylmuramoyl-l-alanyl-d-glutamate:l-lysine ligase was identified and cloned into plasmid vectors. Induction of its overexpression in E. coli rapidly results in abnormal morphological changes and subsequent cell lysis. A reduction of 28% in the peptidoglycan content was observed in induced cells, and analysis of the peptidoglycan composition and structure showed that ca. 50% of themeso-diaminopimelic acid residues were replaced byl-lysine. Lysine was detected in both monomer and dimer fragments, but the acceptor units from the latter contained exclusivelymeso-diaminopimelic acid, suggesting that no transpeptidation could occur between the ɛ-amino group ofl-lysine and the α-carboxyl group ofd-alanine. The overall cross-linking of the macromolecule was only slightly decreased. Detection and analysis ofmeso-diaminopimelic acid- andl-lysine-containing peptidoglycan precursors confirmed the presence of l-lysine in precursors containing amino acids added after the reaction catalyzed by the MurE ligase and provided additional information about the specificity of the enzymes involved in these latter processes.

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Peptidoglycan biosynthesis by preparations from Bacillus licheniformis: cross-linking of newly synthesized chains to preformed cell wall

Biochemical Journal ◽

10.1042/bj1390781 ◽

1974 ◽

Vol 139 (3) ◽

pp. 781-784 ◽

Cited By ~ 28

Author(s):

J. Barrie Ward ◽

Harold R. Perkins

Keyword(s):

Cell Wall ◽

Bacillus Licheniformis ◽

Free Amino ◽

Membrane Preparation ◽

Diaminopimelic Acid ◽

Cross Linking ◽

Amino Group ◽

Peptidoglycan Biosynthesis ◽

Acetyl Group ◽

Degraded Product

A wall-plus-membrane preparation from a Bacillus licheniformis mutant incorporated radioactivity from a peptidoglycan precursor in which the free amino group of diaminopimelic acid was blocked by 14C-labelled acetyl group. This incorporation was penicillin-sensitive. The enzymically degraded product contained cross-linked dimers, showing that newly synthesized peptidoglycan chains had been cross-linked to the pre-existing cell wall.

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Unique Structural Features of the Peptidoglycan of Mycobacterium leprae

Journal of Bacteriology ◽

10.1128/jb.00982-07 ◽

2007 ◽

Vol 190 (2) ◽

pp. 655-661 ◽

Cited By ~ 48

Author(s):

Sebabrata Mahapatra ◽

Dean C. Crick ◽

Michael R. McNeil ◽

Patrick J. Brennan

Keyword(s):

Mycobacterium Leprae ◽

Structural Features ◽

Diaminopimelic Acid ◽

Side Chains ◽

Muramic Acid ◽

Peptidoglycan Biosynthesis ◽

Cross Linkage ◽

Direct Cross ◽

Genome Decay

ABSTRACT The peptidoglycan structure of Mycobacterium spp. has been investigated primarily with the readily cultivable Mycobacterium smegmatis and Mycobacterium tuberculosis and has been shown to contain unusual features, including the occurrence of N-glycolylated, in addition to N-acetylated, muramic acid residues and direct cross-linkage between meso-diaminopimelic acid residues. Based on results from earlier studies, peptidoglycan from in vivo-derived noncultivable Mycobacterium leprae was assumed to possess the basic structural features of peptidoglycans from other mycobacteria, other than the reported replacement of l-alanine by glycine in the peptide side chains. In the present study, we have analyzed the structure of M. leprae peptidoglycan in detail by combined liquid chromatography and mass spectrometry. In contrast to earlier reports, and to the peptidoglycans in M. tuberculosis and M. smegmatis, the muramic acid residues of M. leprae peptidoglycan are exclusively N acetylated. The un-cross-linked peptide side chains of M. leprae consist of tetra- and tripeptides, some of which contain additional glycine residues. Based on these findings and genome comparisons, it can be concluded that the massive genome decay in M. leprae does not markedly affect the peptidoglycan biosynthesis pathway, with the exception of the nonfunctional namH gene responsible for N-glycolylmuramic acid biosynthesis.

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Biochemical Characterization and Physiological Properties of Escherichia coli UDP-N-Acetylmuramate:l-Alanyl-γ-d-Glutamyl-meso- Diaminopimelate Ligase

Journal of Bacteriology ◽

10.1128/jb.00087-07 ◽

2007 ◽

Vol 189 (11) ◽

pp. 3987-3995 ◽

Cited By ~ 27

Author(s):

Mireille Hervé ◽

Audrey Boniface ◽

Stanislav Gobec ◽

Didier Blanot ◽

Dominique Mengin-Lecreulx

Keyword(s):

Escherichia Coli ◽

Biochemical Characterization ◽

De Novo ◽

Diaminopimelic Acid ◽

Kinetic Properties ◽

Gene Encoding ◽

Peptidoglycan Biosynthesis ◽

Physiological Properties

ABSTRACT The UDP-N-acetylmuramate:l-alanyl-γ-d-glutamyl-meso-diaminopimelate ligase (murein peptide ligase [Mpl]) is known to be a recycling enzyme allowing reincorporation into peptidoglycan (murein) of the tripeptide l-alanyl-γ-d-glutamyl-meso-diaminopimelate released during the maturation and constant remodeling of this bacterial cell wall polymer that occur during cell growth and division. Mpl adds this peptide to UDP-N-acetylmuramic acid, thereby providing an economical additional source of UDP-MurNAc-tripeptide available for de novo peptidoglycan biosynthesis. The Mpl enzyme from Escherichia coli was purified to homogeneity as a His-tagged form, and its kinetic properties and parameters were determined. Mpl was found to accept tri-, tetra-, and pentapeptides as substrates in vitro with similar efficiencies, but it accepted the dipeptide l-Ala-d-Glu and l-Ala very poorly. Replacement of meso-diaminopimelic acid by l-Lys resulted in a significant decrease in the catalytic efficacy. The effects of disruption of the E. coli mpl gene and/or the ldcA gene encoding the ld-carboxypeptidase on peptidoglycan metabolism were investigated. The differences in the pools of UDP-MurNAc peptides and of free peptides between the wild-type and mutant strains demonstrated that the recycling activity of Mpl is not restricted to the tripeptide and that tetra- and pentapeptides are also directly reused by this process in vivo. The relatively broad substrate specificity of the Mpl ligase indicates that it is an interesting potential target for antibacterial compounds.

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Cytochemical Studies of Cell Wall Biosynthesis in Staphylococcus Aureus

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100094905 ◽

1972 ◽

Vol 30 ◽

pp. 328-329

Author(s):

Masaatsu Koike ◽

Koichi Nakashima ◽

Kyoko Iida

Keyword(s):

Staphylococcus Aureus ◽

Periodate Oxidation ◽

Early Time ◽

The Other ◽

Penicillin G ◽

Cell Wall Biosynthesis ◽

Septal Wall ◽

Peptidoglycan Biosynthesis ◽

Gram Positive Bacteria ◽

Cross Linkage

Penicillin exerts the activity to inhibit the peptide cross linkage between each polysaccharide backbone at the final stage of wall-peptidoglycan biosynthesis of bacteria. Morphologically, alterations of the septal wall and mesosome in gram-positive bacteria, which were occurred in early time after treatment with penicillin, have been observed. In this experiment, these alterations were cytochemically investigated by means of silver-methenamine staining after periodate oxidation, which is applied for detection of localization of wall mucopolysaccharide.Staphylococcus aureus strain 209P treated with 100 u/ml of penicillin G was divided into two aliquotes. One was fixed by Kellenberger-Ryter's OSO4 fixative at 30, 60 and 120 min after addition of the antibiotic, dehydrated through alcohol series, and embedded in Epon 812 (Specimen A). The other was fixed by 21 glutaraldehyde, dehydrated through glycolmethacrylate series and embedded in glycolmethacrylate mixture, according to Bernhard's method (Specimen B).

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