The First Total Synthesis of Lipid II:  The Final Monomeric Intermediate in Bacterial Cell Wall Biosynthesis

VanNieuwenhze, Michael S.; Mauldin, Scott C.; Zia‐Ebrahimi, Mohammad; Winger, Brian E.; Hornback, William J.; Saha, Shubhayu; Aikins, James; Blaszczak, Larry C.

doi:10.1021/ja017386d

Cited by 118 publications

(97 citation statements)

References 24 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%

See 1 more Smart Citation

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

305

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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“…This enables the C terminus, which consists of the knotted DE-ring system, to form pores in the phospholipid membrane, which ultimately lead to cell leakage and collapse of the vital ion gradients across the membrane. [3] Lipid II [4] is an essential precursor for cell-wall synthesis. [5] It consists of two amino sugars, N-acetyl glucosamine (GlcNAc) and N-acetyl muramic acid (MurNAc), and a pentapeptide (usually H-Ala-d-Glu(Lys-d-Ala-d-Ala-OH)-OH-) that is attached to the carboxyl moiety of MurNAc.…”

Section: Introductionmentioning

confidence: 99%

“…After completion of the reaction, DCM was removed under reduced pressure, and the residue was dissolved in EtOAc (25 mL mmol À1 ). The solution was washed with KHSO 4 (1 n, 3 25 mL), 10 % Na 2 CO 3 (3 25 mL), and brine (1 25 mL), then dried (Na 2 SO 4 ), filtrated, and evaporated in vacuo. The …”

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confidence: 99%

Synthesis of Bicyclic Alkene‐/Alkane‐Bridged Nisin Mimics by Ring‐Closing Metathesis and their Biochemical Evaluation as Lipid II Binders: toward the Design of Potential Novel Antibiotics

et al. 2007

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This report describes the design, synthesis, and biochemical evaluation of alkene‐ and alkane‐bridged AB(C)‐ring mimics of the lantibiotic nisin. Nisin belongs to a class of natural antimicrobial peptides, and has a unique mode of action: its AB(C)‐ring system binds to the pyrophosphate moiety of lipid II. This mode of action was the rationale for the design of smaller nisin‐derived peptides to obtain novel potential antibiotics. As a conformational constraint the thioether bridge was mimicked by an alkene‐ or alkane isostere. The peptides of the linear individual ring precursors were synthesized on solid support or in solution, and cyclized by ring‐closing metathesis in solution with overall yields of between 36 and 89 %. The individual alkene‐bridged macrocycles were assembled in solution by using carbodiimide‐based synthesis protocols for the corresponding AB(C)‐ring mimics. These compounds were tested for their binding affinity toward lipid II by evaluation of their potency to inhibit nisin‐induced carboxyfluorescein release from large unilamellar vesicles. It was found that these AB(C)‐ring mimics were not able to induce membrane leakage; however, they acted by inhibiting nisin‐induced carboxyfluorescein release; this indicates their affinity toward lipid II. These results imply that an alkene or alkane moiety is a suitable thioether bridge mimic.

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The First Total Synthesis of Lipid II: The Final Monomeric Intermediate in Bacterial Cell Wall Biosynthesis

Cited by 118 publications

References 24 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

Synthesis of Bicyclic Alkene‐/Alkane‐Bridged Nisin Mimics by Ring‐Closing Metathesis and their Biochemical Evaluation as Lipid II Binders: toward the Design of Potential Novel Antibiotics

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