Enzymatic Synthesis of Lipid II and Analogues

Huang, Li-Hsin; Huang, Shih-Hsien; Chang, Ya‐Chih; Cheng, Wei‐Chieh; Cheng, Ting‐Jen R.; Wong, Chi‐Huey

doi:10.1002/anie.201402313

Cited by 44 publications

(47 citation statements)

References 38 publications

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“…We recently developed a method to obtain Gly 5 -Lipid II in large amounts from S. aureus , enabling access to Gly 5 -peptidoglycan. 7,8 Here we report that it is possible to obtain substantial quantities of Gly 1 - and Gly 3 -Lipid II from appropriate S. aureus mutant strains, which has enabled us to examine the crosslinking preferences of native S. aureus PBPs as well as PBP2a.…”

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Peptidoglycan Cross-Linking Preferences of Staphylococcus aureus Penicillin-Binding Proteins Have Implications for Treating MRSA Infections

et al. 2017

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Methicillin-resistant Staphylococcus aureus (MRSA) infections are a global public health problem. MRSA strains have acquired a non-native penicillin-binding protein called PBP2a that crosslinks peptidoglycan when the native S. aureus PBPs are inhibited by β-lactams. If assembly of the pentaglycine branch on the cell wall precursor Lipid II is genetically blocked, MRSA strains become susceptible to β-lactams. Therefore, it has been proposed that PBP2a can only crosslink peptidoglycan strands bearing a complete pentaglycine branch. This hypothesis has never been tested because the necessary substrates have not been available. Here, we obtained S. aureus Lipid II variants having shorter glycine branches and have tested whether PBP2a and two other S. aureus transpeptidases, PBP2 and PBP4, can crosslink peptidoglycan strands made from the variants. There are striking differences in enzymatic activity among these enzymes depending on the length of the glycine branch, but we find that PBP2a can, in fact, crosslink glycan strands bearing triglycine. We report experiments in cells that are consistent with our in vitro findings about the crosslinking preferences of these PBPs. In addition to providing insights into the cell wall physiology of a major pathogen, our studies identify the best target for β-lactam potentiators to treat MRSA.

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Peptidoglycan Cross-Linking Preferences of Staphylococcus aureus Penicillin-Binding Proteins Have Implications for Treating MRSA Infections

et al. 2017

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“…Chemical, chemoenzymatic, and biosynthetic routes to Lipid II variants have been developed, 13,14,18–27 but all are laborious. Moreover, each approach was developed for a specific Lipid II variant and considerable reengineering of the routes is required to obtain other Lipid II variants.…”

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Lipid II overproduction allows direct assay of transpeptidase inhibition by β-lactams

et al. 2017

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Peptidoglycan is an essential crosslinked polymer that surrounds bacteria and protects them from osmotic lysis. Beta-lactam antibiotics target the final stages of peptidoglycan biosynthesis by inhibiting the transpeptidases that crosslink glycan strands to complete cell wall assembly. Characterization of transpeptidases and their inhibition by beta-lactams has been hampered by lack of access to substrate. We describe a general approach to accumulate Lipid II in bacteria and to obtain large quantities of this cell wall precursor. We demonstrate utility by isolating Staphylococcus aureus Lipid II and reconstituting the synthesis of crosslinked peptidoglycan by the essential penicillin-binding protein 2, PBP2, which catalyzes both glycan polymerization and transpeptidation. We also show that we can compare the potencies of different beta-lactams by directly monitoring transpeptidase inhibition. The methods reported here will enable a better understanding of cell wall biosynthesis and facilitate studies of next-generation transpeptidase inhibitors.

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“…The enzymatic approach utilises MraY and MurG activities of membrane preparations to synthesise lipid II in the presence of undecaprenyl phosphate, UDP‐MurNAC‐pentapeptide and UDP‐GlcNAc. As both enzymes have narrow substrate specificity, this method is highly effective and can be easily scaled up . On the other hand, chemical synthesis enables various structural alterations of lipid II that cannot be achieved with MraY and MurG .…”

Section: Targeting the Central Component Of Peptidoglycan Synthesismentioning

confidence: 99%

New Found Hope for Antibiotic Discovery: Lipid II Inhibitors

Chan

2016

Chemistry A European J

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Abstract:Research into antibacterial agents has recently gathered pace in light of the disturbing crisis of antimicrobial resistance. Development of modern tools offers opportunity of reviving the fallen era of antibacterial discovery through uncovering novel lead compounds that target vital bacterial cell components, such as lipid II. This paper provides a summary of the role of lipid II as well as an overview and insight into the structural features of macrocyclic peptides that inhibit this bacterial cell wall component. The recent discovery of teixobactin, a new class of lipid II inhibitor has generated substantial research interests. As such, the significant progress that has been achieved towards its development as a promising antibacterial agent is discussed.

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Enzymatic Synthesis of Lipid II and Analogues

Cited by 44 publications

References 38 publications

Peptidoglycan Cross-Linking Preferences of Staphylococcus aureus Penicillin-Binding Proteins Have Implications for Treating MRSA Infections

Peptidoglycan Cross-Linking Preferences of Staphylococcus aureus Penicillin-Binding Proteins Have Implications for Treating MRSA Infections

Lipid II overproduction allows direct assay of transpeptidase inhibition by β-lactams

New Found Hope for Antibiotic Discovery: Lipid II Inhibitors

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