Forming crosslinked peptidoglycan from synthetic Gram-negative Lipid II

The bacterial cell wall precursor, Lipid II, has a highly conserved structure among different organisms except for differences in the amino acid sequence of the peptide side chain. Here, we report an efficient and flexible synthesis of the canonical Lipid II precursor required for the assembly of Gr...

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Bibliographic Details
Published inJournal of the American Chemical Society Vol. 135; no. 12; pp. 4632 - 4635
Main Authors Lebar, Matthew D., Lupoli, Tania J., Tsukamoto, Hirokazu, May, Janine M., Walker, Suzanne, Kahne, Daniel
Format Journal Article
LanguageEnglish
Published 13.03.2013
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Summary:The bacterial cell wall precursor, Lipid II, has a highly conserved structure among different organisms except for differences in the amino acid sequence of the peptide side chain. Here, we report an efficient and flexible synthesis of the canonical Lipid II precursor required for the assembly of Gram-negative peptidoglycan (PG). We use a rapid LC/MS assay to analyze PG glycosyltransfer (PGT) and transpeptidase (TP) activities of Escherichia coli penicillin binding proteins PBP1A and PBP1B and show that the native m -DAP residue in the peptide side chain of Lipid II is required in order for TP-catalyzed peptide crosslinking to occur in vitro . Comparison of PG produced from synthetic canonical E. coli Lipid II with PG isolated from E. coli cells demonstrates that we can produce PG in vitro that resembles native structure. This work provides the tools necessary for reconstituting cell wall synthesis, an essential cellular process and major antibiotic target, in a purified system.
Bibliography:Present Addresses
Graduate School of Pharmaceutical Sciences and Pharmaceutical Chemistry, Tohoku University, Sendai, Miyagi, Japan
ISSN:0002-7863
1520-5126
DOI:10.1021/ja312510m