Presence of UDP-N-acetylmuramyl-hexapeptides and -heptapeptides in enterococci and staphylococci after treatment with ramoplanin, tunicamycin, or vancomycin

Billot-Klein, D; Shlaes, David M.; Bryant, D T W; Bell, D; Legrand, Raymond; Gutmann, Laurent; Heijenoort, Jean van

doi:10.1128/jb.179.15.4684-4688.1997

Cited by 46 publications

(34 citation statements)

References 33 publications

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“…faecium strain MT10 Rev (Billot-Klein et al, 1997), or Bacillus cereus ATCC 11778 was grown aerobically in a 10 l fermenter in BHI\CAA medium. The substrate UDPMurNAc-pentapeptide, containing either -lysine or mesodiaminopimelic acid, was isolated from these cells as described (Kohlrausch & Holtje, 1991b), with minor modifications.…”

Section: Purification Of Udp-murnac-pentapeptide From Bacteriamentioning

confidence: 99%

Antibacterial activity of synthetic analogues based on the disaccharide structure of moenomycin, an inhibitor of bacterial transglycosylase

et al. 2000

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Moenomycin is a natural product glycolipid that inhibits the growth of a broad spectrum of Gram-positive bacteria. In Escherichia coli, moenomycin inhibits peptidoglycan synthesis at the transglycosylation stage, causes accumulation of cell-wall intermediates, and leads to lysis and cell death. However, unlike Esc. coli, where 5-6 log units of killing are observed, 0-2 log units of killing occurred when Gram-positive bacteria were treated with similar multiples of the MIC. In addition, bulk peptidoglycan synthesis in intact Gram-positive cells was resistant to the effects of moenomycin. In contrast, synthetic disaccharides based on the moenomycin disaccharide core structure were identified that were bactericidal to Gram-positive bacteria, inhibited cell-wall synthesis in intact cells, and were active on both sensitive and vancomycinresistant enterococci. These disaccharide analogues do not inhibit the formation of N-acetylglucosamine-β-1,4-MurNAc-pentapeptide-pyrophosphorylundecaprenol (lipid II), but do inhibit the polymerization of lipid II into peptidoglycan in Esc. coli. In addition, cell growth was required for bactericidal activity. The data indicate that synthetic disaccharide analogues of moenomycin inhibit cell-wall synthesis at the transglycosylation stage, and that their activity on Gram-positive bacteria differs from moenomycin due to differential targeting of the transglycosylation process. Inhibition of the transglycosylation process represents a promising approach to the design of new antibacterial agents active on drug-resistant bacteria.

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Section: Purification Of Udp-murnac-pentapeptide From Bacteriamentioning

confidence: 99%

Antibacterial activity of synthetic analogues based on the disaccharide structure of moenomycin, an inhibitor of bacterial transglycosylase

et al. 2000

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“…The assay for UDP-MurNAc-hexapeptide synthesis proved useful in identifying the E. faecalis ligase (Table 1), although this enzyme is likely to preferentially act in vivo on the lipid intermediates, as reported for several membrane-associated ligases of gram-positive cocci (enterococci, staphylococci, and streptococci) (3,8,14). W. viridescens is an exception since it produces a soluble ligase for addition of L-alanine to cytoplasmic UDP-MurNAc-pentapeptide (8,15).…”

Section: Discussionmentioning

confidence: 99%

“…This conclusion is supported by the detection of only small amounts (Ͻ4%) of UDP-MurNAc-hexapeptide in Staphylococcus haemolyticus and S. aureus in spite of high-level accumulation (ca. 12-fold) of UDP-MurNAc-pentapeptide in bacteria treated with ramoplanin, which blocks lipid II synthesis (3). Similarly, moderate accumulation of L-Ala-containing UDP-MurNAc-hexapeptide was detected in penicillin-treated Enterococcus faecalis (12).…”

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confidence: 95%

Identification of the UDP-MurNAc-Pentapeptide: l -Alanine Ligase for Synthesis of Branched Peptidoglycan Precursors in Enterococcus faecalis

et al. 2001

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Many species of gram-positive bacteria produce branched peptidoglycan precursors resulting from the transfer of various L-amino acids or glycine from amino acyl-tRNA to the -amino group of L-lysine. The UDP-MurNAc-pentapeptide:L-alanine ligase and alanyl-tRNA synthetase genes from Enterococcus faecalis were identified, cloned, and overexpressed in Escherichia coli. The purified enzymes were necessary and sufficient for tRNA-dependent addition of L-alanine to UDP-MurNAc-pentapeptide in vitro. The ligase belonged to the Fem family of proteins, which were initially identified genetically as factors essential for methicillin resistance in Staphylococcus aureus.In gram-positive bacteria, the ε-amino group of L-lysine in the pentapeptide stem of peptidoglycan precursors is often replaced by amino acid chains of various lengths and compositions (19) (Fig. 1). These amino acids form cross bridges between L-Lys 3 and D-Ala 4 following cross-linking of the peptide stems from nascent chains of peptidoglycan by the D,Dtranspeptidases. The ligases for the addition of glycine and L-amino acids to the ε-amino group of L-lysine use aminoacyltRNAs as substrates, whereas D-amino acids are added in a tRNA-independent reaction (15,20).In Staphylococcus aureus, the femA, femB, and fmhB genes were shown to be essential for incorporation of glycine into the side chain of peptidoglycan precursors (18, 21). The femAB locus was initially identified as a factor essential for methicillin resistance (fem) based on random insertional inactivation of chromosomal genes and a screen for reduced expression of resistance mediated by the penicillin binding protein 2A (PBP2A) (2, 5). Inactivation of femA or femB was subsequently reported to prevent incorporation of glycine residues at positions 2 to 5 or positions 4 to 5 of the penta-glycine cross bridge since muropeptides cross-linked by one or three glycine residues were detected in the corresponding mutants (4, 21). Inactivation of fmhB, formerly femX, is lethal, but the construction of a mutant conditionally expressing fmhB under the control of a xylose-inducible promoter showed that the gene was essential for synthesis of branched peptidoglycan precursors (18). These results indicated that the fem gene products were required for incorporation of glycine at positions 1 (FmhB), 2 and 3 (FemA), and 4 and 5 (FemB) of the cross bridge, although the catalytic activity of the proteins was not directly assessed (18). Similarly, inactivation of two fmhB homologues in Streptococcus pneumoniae, designated murM (fibA) and murN (fibB), reduced addition of L-Ala or L-Ser to the ε-amino group of L-Lys and subsequent addition of a second L-Ala residue, respectively (6, 7, 23). Overall, disruption of the murMN operon reduced the proportion of branched peptide stems in the peptidoglycan from 89 to 33% (6). In contrast to what occurs in S. aureus (18), direct cross-linking of L-Lys to D-Ala occurs in S. pneumoniae, and the murMN operon was accordingly reported to be unessential (6). However, production of branch...

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“…The peptidyltransferases from Enterococcus faecalis (BppA1 and BppA2) and Enterococcus faecium (Aslfm) were overproduced and purified by using UDP-MurNAc-pentapeptide as an in vitro substrate (15,23,24). However, since the turnover observed with this substrate was very low and since no large UDP-MurNAc-hexapeptide pool level has been encountered in studies of enterococci (20), it can be assumed that in vivo the addition of the bridging amino acids occurs on the lipid intermediates. It remains to be determined whether this takes place on lipid I, on lipid II, or on both.…”

Section: Biosynthesis Of Modified Lipid Intermediatesmentioning

confidence: 99%

Lipid Intermediates in the Biosynthesis of Bacterial Peptidoglycan

Heijenoort

2007

Microbiol Mol Biol Rev

172

135

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SUMMARY This review is an attempt to bring together and critically evaluate the now-abundant but dispersed data concerning the lipid intermediates of the biosynthesis of bacterial peptidoglycan. Lipid I, lipid II, and their modified forms play a key role not only as the specific link between the intracellular synthesis of the peptidoglycan monomer unit and the extracytoplasmic polymerization reactions but also in the attachment of proteins to the bacterial cell wall and in the mechanisms of action of antibiotics with which they form specific complexes. The survey deals first with their detection, purification, structure, and preparation by chemical and enzymatic methods. The recent important advances in the study of transferases MraY and MurG, responsible for the formation of lipids I and II, are reported. Various modifications undergone by lipids I and II are described, especially those occurring in gram-positive organisms. The following section concerns the cellular location of the lipid intermediates and the translocation of lipid II across the cytoplasmic membrane. The great efforts made since 2000 in the study of the glycosyltransferases catalyzing the glycan chain formation with lipid II or analogues are analyzed in detail. Finally, examples of antibiotics forming complexes with the lipid intermediates are presented.

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Presence of UDP-N-acetylmuramyl-hexapeptides and -heptapeptides in enterococci and staphylococci after treatment with ramoplanin, tunicamycin, or vancomycin

Cited by 46 publications

References 33 publications

Antibacterial activity of synthetic analogues based on the disaccharide structure of moenomycin, an inhibitor of bacterial transglycosylase

Antibacterial activity of synthetic analogues based on the disaccharide structure of moenomycin, an inhibitor of bacterial transglycosylase

Identification of the UDP-MurNAc-Pentapeptide: l -Alanine Ligase for Synthesis of Branched Peptidoglycan Precursors in Enterococcus faecalis

Lipid Intermediates in the Biosynthesis of Bacterial Peptidoglycan

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