Specific Binding of Nisin to the Peptidoglycan Precursor Lipid II Combines Pore Formation and Inhibition of Cell Wall Biosynthesis for Potent Antibiotic Activity*

By I. Wiedemann, E. Breukink, C. van Kraaij, Oscar P. Kuipers, G. Bierbaum, B. de Kruijff, H. Sahl. Journal of Biological Chemistry, 2001

739 citations recorded by the source corpus.

Unlike numerous pore-forming amphiphilic peptide antibiotics, the lantibiotic nisin is active in nanomolar concentrations, which results from its ability to use the lipid-bound cell wall precursor lipid II as a docking molecule for subsequent pore formation. Here we use genetically engineered nisin variants to identify the structural requirements for the interaction of the peptide with lipid II. Mutations affecting the conformation of the N-terminal part of nisin comprising rings A through C, e.g.[S3T]nisin, led to reduced binding and increased the peptide concentration necessary for pore formation. The binding constant for the S3T mutant was 0.043 × 107 m −1 compared with 2 × 107 m −1 for the wild-type peptide, and the minimum concentration for pore formation increased from the 1 nm to the 50 nm range. In contrast, peptides mutated in the flexible hinge region, e.g. [ΔN20/ΔM21]nisin, were completely inactive in the pore formation assay, but were reduced to some extent in their in vivo activity. We found the remaining in vivo activity to result from the unaltered capacity of the mutated peptide to bind to lipid II and thus to inhibit its incorporation into the peptidoglycan network. Therefore, through interaction with the membrane-bound cell wall precursor lipid II, nisin inhibits peptidoglycan synthesis and forms highly specific pores. The combination of two killing mechanisms in one molecule potentiates antibiotic activity and results in nanomolar MIC values, a strategy that may well be worth considering for the construction of novel antibiotics.

Subfields: Economics and Econometrics, Finance

View Paper: https://www.semanticscholar.org/paper/ed9cbd6686324084572374902d61c5798d2e126a

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1. M. Øverland
2. L. Marraffini
3. Dina Raafat
4. B. Kruijff
5. Andrea C. DeDent
6. O. Schneewind
7. Lucy H. Deegan
8. F. Reveco

Citations in the corpus

• Preservation and fermentation: past, present and future. (2002, background): “Indeed, in the absence of this precursor, significantly higher concentrations of nisin are required to form pores (Breukink et al., 1999; Wiedemann et al., 2001).”
• Insights into the Mode of Action of Chitosan as an Antibacterial Compound (2008, background): “Nisin is known to form a defined and stable pore, using the membrane-bound bacterial cell wall precursor lipid II as an anchor molecule, simultaneously resulting in inhibition of cell wall biosynthesis (50, 51).”
• Bacteriocins: Biological tools for bio-preservation and shelf-life extension (2006, background): “In addition to pore formation, it can also inhibit cell wall biosynthesis through the binding of lipid II—a step which in fact probably precedes pore formation (Breukink et al., 1999; Weidemann et al., 2001).”
• Sortases and the Art of Anchoring Proteins to the Envelopes of Gram-Positive Bacteria (2006, methodology): “The C-terminal anchor of immunoprecipitated (32)P-labeled surface protein was analyzed by thin-layer chromatography and observed to bind nisin (154), an antibiotic that specifically interacts with lipid II (214).”
• Intestinal bacterial community structure differs between healthy and inflamed intestines in Atlantic salmon (Salmo salar L.) (2014, background): “lactis in the SBM group can be related to its capability of producing antimicrobial peptides and proteins that have activity against some gram-positive and negative bacteria (Twomey et al., 2002; Wiedemann et al., 2001), and thus modulating the intestinal microbiota by antagonizing certain bacteria ”
• Lipid II as a target for antibiotics (2006, positive): “Support for this comes from the observation that in the presence of Lipid II, nisin activity was not dependent on the membrane lipid composition with respect to anionic lipid content over a broad concentration range 25 .”