Incorporation of LL-diaminopimelic acid into peptidoglycan of Escherichia coli mutants lacking diaminopimelate epimerase encoded by dapF

Mengin‐Lecreulx, Dominique; Michaud, Catherine; Richaud, Catherine; Blanot, Didier; Heijenoort, Jean van

doi:10.1128/jb.170.5.2031-2039.1988

Cited by 61 publications

(70 citation statements)

References 32 publications

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“…Moreover, tripeptide lipid II was used as a substrate in the in vitro assay for PBP lb polymerase activity apparently as efficiently as was pentapeptide lipid II. A number of observations indicate that in vivo the E. coli peptidoglycan biosynthesis system will accept modifications in its peptide moiety (24).…”

Section: Resultsmentioning

confidence: 99%

Membrane intermediates in the peptidoglycan metabolism of Escherichia coli: possible roles of PBP 1b and PBP 3

et al. 1992

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The two membrane precursors (pentapeptide lipids I and II) of peptidoglycan are present in Escherichia coli at cell copy numbers no higher than 700 and 2,000 respectively. Conditions were determined for an optimal accumulation of pentapeptide lipid II from UDP-MurNAc-pentapeptide in a cell-free system and for its isolation and purification. When UDP-MurNAc-tripeptide was used in the accumulation reaction, tripeptide lipid II was formed, and it was isolated and purified. Both lipids II were compared as substrates in the in vitro polymerization by transglycosylation assayed with PBP lb or PBP 3. With PBP lb, tripeptide lipid II was used as efficiently as pentapeptide lipid II. It should be stressed that the in vitro PBP lb activity accounts for at best to 2 to 3% of the in vivo synthesis. With PBP 3, no polymerization was observed with either substrate. Furthermore, tripeptide lipid II was detected in D-cycloserine-treated cells, and its possible in vivo use in peptidoglycan formation is discussed. In particular, it is speculated that the transglycosylase activity of PBP lb could be coupled with the transpeptidase activity of PBP 3, using mainly tripeptide lipid II as precursor.It is now established that in Escherichia coli there are at least two distinct modes for the insertion of newly polymerized peptidoglycan material, one for elongation and the other for septation (30). Several proteins have been described as directly involved in the polymerization reactions (34, 47). In particular, certain penicillin-binding proteins (PBPs) were found to catalyze in vitro polymerization reactions with Nacetylglucosaminyl -N-acetylmuramyl(pentapeptide) -pyrophosphate-undecaprenol as a substrate (17,18,22,29,39,41). In vivo, this membrane intermediate (lipid II) is formed from UDP-N-acetylmuramoyl-pentapeptide by a two step process (34). First, a translocase catalyzes the transfer of the phospho-MurNAc-pentapeptide moiety of UDP-MurNAcpentapeptide to the membrane acceptor undecaprenol phosphate, yielding MurNAc(pentapeptide)-pyrophosphate-undecaprenol-(lipid I). Thereafter, a transferase catalyzes the addition of N-acetylglucosamine, yielding lipid II. The translocase also catalyzes the reverse reaction, and the equilibrium is in favor of the formation of UDP-MurNAc-pentapeptide (10,26 cell-free system in which it was allowed to accumulate to a certain extent by incubation of membranes with UDPGlcNAc and radiolabelled UDP-MurNAc-pentapeptide (44).In this study, a set of optimal conditions for a maximal in vitro formation of lipid II with particulate fraction from E. coli was sought, as well as substantial improvements in its purification. A lipid II containing tripeptide Ala-y-D-Glumeso-DAP instead of the pentapeptide was prepared in a similar way. Its possible in vivo presence was investigated. Both types of lipid II were compared as substrates in the in vitro polymerization reactions assayed with PBP lb or PBP 3. The possible physiological significance of such reactions are discussed. MATERUILS AND METHODSBacterial strains, ...

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Section: Resultsmentioning

confidence: 99%

Membrane intermediates in the peptidoglycan metabolism of Escherichia coli: possible roles of PBP 1b and PBP 3

et al. 1992

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“…0.04, 36 and 1.5 mM, respectively [15,17]. MurE did not follow simple kinetics with the fairly good substrate 3a: v vs. log[S] plots were bell-shaped with the top near 10 mM (not shown).…”

Section: Fluorinated Analogue 8 [(2s3r6s)-3-fluoro-a2pmmentioning

confidence: 96%

“…1) to assay them as substrates or inhibitors of MurE from Escherichia coli. [17] and meso-A2pm [18] were obtained according to published procedures, meso-[laC]A2pm (11.6 TBq mo1-1) was purchased from CEA (Saclay, France). The synthesis of A2pm analogues 2-8 and 10 has already been described [5,6,8,9,11]; analogue 9 was synthesized according to the procedure of Girodeau et al [10].…”

Section: Introductionmentioning

confidence: 99%

Effect of analogues of diaminopimelic acid on the meso‐diaminopimelate‐adding enzyme from Escherichia coli

et al. 1996

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Several analogues of diaminopimelic acid (A2pm) were tested as substrates or inhibitors of the meso-diaminopimelate-adding enzyme from Escherichia coll. They included lanthionine derivatives, a phosphonic analogue, heterocyclic compounds, 3-fiuoro-A2pm, 4-methylene-A2pm and N-hydroxyA2pm. The best substrates were, in decreasing order of specific enzyme activity, (2S,3R,6S)-3-fluoro-A2pm, meso-lanthioulne sulfoxide and N-hydroxy-A2pm (mixture of stereoisomers). In those cases where all the stereoisomers were available, the specificity could be described as meso >> DD = LL. N-HydroxyA2pm (mixture of stereoisomers) strongly inhibited the addition of radioactive meso-A2pm to UDP-N-acetylmuramoyl-dipeptide.

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“…The chemical synthesis of the meso-lanthionine and L-allocystathionine has been described previously (10). The purity of the DAP, lanthionine, and cystathionine isomers could be assessed using a high performance liquid chromatography (HPLC) technique after derivatization with the Marfey reagent (11,12). meso-DAP (D)-monoamide (DAP amide ) dihydrobromide was prepared by treatment of N,NЈ-bisbenzyloxycarbonyl-meso-DAP (D)-monoamide (13) with HBr/acetic acid for 1 h.…”

Section: Methodsmentioning

confidence: 99%

Peptidoglycan Molecular Requirements Allowing Detection by Nod1 and Nod2

Girardin

Travassos

Hervé

et al. 2003

Journal of Biological Chemistry

613

602

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Nod1 and Nod2 are mammalian proteins implicated in the intracellular detection of pathogen-associated molecular patterns. Recently, naturally occurring peptidoglycan (PG) fragments were identified as the microbial motifs sensed by Nod1 and Nod2. Whereas Nod2 detects GlcNAc-MurNAc dipeptide (GM-Di), Nod1 senses a unique diaminopimelate-containing GlcNAc-MurNAc tripeptide muropeptide (GM-Tri DAP ) found mostly in Gram-negative bacterial PGs. Because Nod1 and Nod2 detect similar yet distinct muropeptides, we further analyzed the molecular sensing specificity of Nod1 and Nod2 toward PG fragments. Using a wide array of natural or modified muramyl peptides, we show here that Nod1 and Nod2 have evolved divergent strategies to achieve PG sensing. By defining the PG structural requirements for Nod1 and Nod2 sensing, this study reveals how PG processing and modifications, either by host or bacterial enzymes, may affect innate immune responses. Peptidoglycan (PG)1 is a major constituent of the cell wall of Gram-positive bacteria and consists of glycan chains of alternating N-acetylglucosamine (GlcNAc) and N-acetylmuramic acid (MurNAc) that are cross-linked to each other by short peptides, allowing the formation of a rigid polymer that surrounds the bacterial cell. During bacterial life cycle, peptidoglycan is constantly degraded by specific hydrolases, and newly synthesized subunits are integrated into the polymeric structure, allowing biological processes such as cell division. In Gram-negative bacteria, a thin layer of peptidoglycan is also found in the periplasmic space. Apart from the thickness and the degree of stem peptides cross-linking, an important difference between Gram-positive and Gram-negative peptidoglycans resides in the nature of the third amino acid of the peptides. In Gram-positive bacteria, this amino acid is commonly a lysine, whereas a diaminopimelic acid is found in most Gramnegative bacteria. Extensive analysis from many bacteria has revealed that, in steady state conditions, peptidoglycans from each bacterial strain have somehow fixed proportions of di-, tri-, tetra-, or pentapeptide substituted to the MurNAc sugar moiety.

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Incorporation of LL-diaminopimelic acid into peptidoglycan of Escherichia coli mutants lacking diaminopimelate epimerase encoded by dapF

Cited by 61 publications

References 32 publications

Membrane intermediates in the peptidoglycan metabolism of Escherichia coli: possible roles of PBP 1b and PBP 3

Membrane intermediates in the peptidoglycan metabolism of Escherichia coli: possible roles of PBP 1b and PBP 3

Effect of analogues of diaminopimelic acid on the meso‐diaminopimelate‐adding enzyme from Escherichia coli

Peptidoglycan Molecular Requirements Allowing Detection by Nod1 and Nod2

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