Lipid II:  Total Synthesis of the Bacterial Cell Wall Precursor and Utilization as a Substrate for Glycosyltransfer and Transpeptidation by Penicillin Binding Protein (PBP) 1b of Eschericia coli

Schwartz, Benjamin; Markwalder, Jay A.; Wang, Yi

doi:10.1021/ja0166848

Cited by 130 publications

(132 citation statements)

References 19 publications

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“…Furthermore, lipid II contains a 55-carbon polyprenyl chain that renders it insoluble in water and difficult to handle in enzyme assays. To enable the study of bacterial transglycosylases, we and others (10,20,21) have established synthetic routes to natural lipid II and analogues containing truncated polyprenyl chains (10). We previously reported that compound 10, which contains a 35-carbon lipid chain, is a better substrate than natural lipid II to use for monitoring PBP1b activity in vitro (10).…”

Section: Resultsmentioning

confidence: 99%

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Vancomycin analogues active against vanA-resistant strains inhibit bacterial transglycosylase without binding substrate

Chen

Walker

Sun

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

139

147

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Bacterial transglycosylases are enzymes that couple the disaccharide subunits of peptidoglycan to form long carbohydrate chains. These enzymes are the target of the pentasaccharide antibiotic moenomycin as well as the proposed target of certain glycopeptides that overcome vancomycin resistance. Because bacterial transglycosylases are difficult enzymes to study, it has not previously been possible to evaluate how moenomycin inhibits them or to determine whether glycopeptide analogues directly target them. We have identified transglycosylase assay conditions that enable kinetic analysis of inhibitors and have examined the inhibition of Escherichia coli penicillin-binding protein 1b (PBP1b) by moenomycin as well as by various glycopeptides. We report that chlorobiphenyl vancomycin analogues that are incapable of binding substrates nevertheless inhibit E. coli PBP1b, which shows that these compounds interact directly with the enzyme. These findings support the hypothesis that chlorobiphenyl vancomycin derivatives overcome vanA resistance by targeting bacterial transglycosylases. We have also found that moenomycin is not competitive with respect to the lipid II substrate of PBP1b, as has long been believed. With the development of suitable methods to evaluate bacterial transglycosylases, it is now possible to probe the mechanism of action of some potentially very important antibiotics.

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Section: Resultsmentioning

confidence: 99%

“…A major impediment to studying these enzymes has been that adequate supplies of lipid II cannot be obtained from natural sources. The development of synthetic approaches to make lipid II and analogues has solved the substrate problem (10,20,21), making it possible to assay inhibitors.…”

Section: Discussionmentioning

confidence: 99%

Vancomycin analogues active against vanA-resistant strains inhibit bacterial transglycosylase without binding substrate

Chen

Walker

Sun

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

139

147

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“…So far, access to Lipid II and variants was limited to groups having considerable synthetic expertise (14,16,17). We reasoned that it should be possible to produce Lipid II if membrane preparations carrying sufficient MraY and MurG activity were provided with the appropriate UDP-activated amino sugars and undecaprenyl phosphate in the presence of a suitable detergent.…”

Section: Synthesis Of Lipid II Andmentioning

confidence: 99%

Lipid II Is an Intrinsic Component of the Pore Induced by Nisin in Bacterial Membranes

Breukink

Heusden

Vollmerhaus

et al. 2003

Journal of Biological Chemistry

303

311

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The peptidoglycan layers surrounding bacterial membranes are essential for bacterial cell survival and provide an important target for antibiotics. Many antibiotics have mechanisms of action that involve binding to Lipid II, the prenyl chain-linked donor of the peptidoglycan building blocks. One of these antibiotics, the pore-forming peptide nisin uses Lipid II as a receptor molecule to increase its antimicrobial efficacy dramatically. Nisin is the first example of a targeted membranepermeabilizing peptide antibiotic. However, it was not known whether Lipid II functions only as a receptor to recruit nisin to bacterial membranes, thus increasing its specificity for bacterial cells, or whether it also plays a role in pore formation. We have developed a new method to produce large amounts of Lipid II and variants thereof so that we can address the role of the lipidlinked disaccharide in the activity of nisin. We show here that Lipid II is not only the receptor for nisin but an intrinsic component of the pore formed by nisin, and we present a new model for the pore complex that includes Lipid II.The cell wall is an essential structure of a bacterium, providing its shape and protecting it from bursting because of the high osmotic pressures of the cytoplasm. This wall is a threedimensional network built of identical subunits consisting of two amino sugars, N-acetylglucosamine (GlcNAc) and N-acetylmu-is attached to the carboxyl group of MurNAc. These subunits are assembled in the cytosol of bacteria using UDP-activated precursors on a special lipid carrier, undecaprenyl phosphate (for a review see Ref. 1). The integral membrane protein MraY and the peripherally membraneassociated MurG that synthesize the precursors Lipid I and II, respectively, are the key enzymes in the last two cytoplasmic steps in the formation of the subunits (Fig. 1). Subsequently, Lipid II is transported across the plasma membrane via an as of yet unknown mechanism. Thereafter, the subunits are polymerized and inserted into the pre-existing cell wall by means of the penicillin-binding proteins (for review see Ref.2). Numerous antibiotics target the cell wall synthesis, including a diverse group of antibiotics that bind to Lipid II. Perhaps the best known of these antibiotics is vancomycin, the antibiotic of last resort to treat MRSA infections (3). However, there are many others, including the polypeptide nisin, that kill cells by permeabilizing bacterial membranes. Efficient membrane permeabilization by nisin requires an interaction with Lipid II (4, 5). This designates nisin as the first example of a targeted poreforming peptide antibiotic.Two recent studies have shed light on the structural requirements within nisin for the interaction with Lipid II. A genetic approach indicated that the N terminus of nisin is involved in the interaction with Lipid II (6), and more recently we could map the binding interface toward specific residues in the N terminus using 15 N-labeled nisin (7). It is not clear yet what events lead to pore formation after the...

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“…The cell walls of all bacteria contain a layer of peptidoglycan, a biopolymer of the alternating amino sugars N-acetylglucosamine (D-NAc-Glu, NAG) and N-acetylmuramic acid (NAc-Mur, NAM), which is modified with pentapeptides of the sequence L-alanyl--D-glutamyl-L-lysyl-D-alanyl-D-alanine (in the case of S. aureus) that is attached to the 3-hydroxy group of the NAc-Mur sugar (Schwartz et al 2001;Breukink and de Kruijff 2006;de Kruijff et al 2008). The cross linkage of the peptides by penicillin binding proteins (PBPs) to macromolecules confers a structural rigidity and mechanical strength that prevents the bacterial protoplast to burst under the osmotic internal pressure.…”

Section: Lessons Learned From Druggable Targets In Bacteriamentioning

confidence: 99%

New Structural Templates for Clinically Validated and Novel Targets in Antimicrobial Drug Research and Development

Klahn

Brönstrup

2016

Current Topics in Microbiology and Immunology

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Abstract:The development of bacterial resistance against current antibiotic drugs necessitates a continuous renewal of the arsenal of efficacious drugs. This imperative has not been met by the output of antibiotic research and development of the past decades for various reasons, including the declining efforts of large pharma companies in this area. Moreover, the majority of novel antibiotics are chemical derivatives of existing structures that represent mostly step innovations, implying that the available chemical space may be exhausted. This review negates this impression by showcasing recent achievements in lead finding and optimization of antibiotics that have novel or unexplored chemical structures. Not surprisingly, many of the novel structural templates like teixobactins, lysocin, griselimycin, or the albicidin/cystobactamid pair were discovered from natural sources. Additional compounds were obtained from the screening of synthetic libraries and chemical synthesis, including the gyrase-inhibiting NTBI s a d spiropyrimidinetrione, the tarocin and targocil inhibitors of wall teichoic acid synthesis, or the boronates and diazabicyclo[3.2

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Lipid II: Total Synthesis of the Bacterial Cell Wall Precursor and Utilization as a Substrate for Glycosyltransfer and Transpeptidation by Penicillin Binding Protein (PBP) 1b of Eschericia coli

Cited by 130 publications

References 19 publications

Vancomycin analogues active against vanA-resistant strains inhibit bacterial transglycosylase without binding substrate

Vancomycin analogues active against vanA-resistant strains inhibit bacterial transglycosylase without binding substrate

Lipid II Is an Intrinsic Component of the Pore Induced by Nisin in Bacterial Membranes

New Structural Templates for Clinically Validated and Novel Targets in Antimicrobial Drug Research and Development

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