Pore Formation by Nisin Involves Translocation of Its C-Terminal Part across the Membrane

Kraaij, Cindy van; Breukink, Eefjan; Noordermeer, Minke A.; Demel, R.A.; Siezen, Roland J.; Kuipers, Oscar P.; Kruijff, Ben de

doi:10.1021/bi980931b

Cited by 77 publications

(74 citation statements)

References 28 publications

(59 reference statements)

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“…The high potency that derives from such a target-mediated poration may well explain why nisin is so much more effective against bacteria in which the docking molecule is available for binding as compared with a peptide that apparently acts in a nontargeted fashion such as magainin 2 (5). The results reported here and in previous papers on the molecular mechanisms of activity of nisin in the absence of lipid II (7,8,18,24,28,36) show that nisin can permeabilize membranes via two different mechanisms. At high concentrations (M range) pores can be formed without lipid II, i.e.…”

Section: Discussionsupporting

confidence: 73%

“…The N-terminal segment of the lantibiotic is essential for binding, and a negative surface charge is not necessary. The C-terminal part of nisin is then assumed to translocate across the membrane, in accordance with the translocation found in the absence of lipid II (28). For this step the flexible hinge region between ring clusters A, B, C and D, E is important.…”

Section: Figmentioning

confidence: 71%

“…only 50% of the incorporated lipid II molecules should be outwardly orientated. However, recent experiments with C-terminal His-tagged nisin Z convincingly demonstrate that the lantibiotic is able to translocate across a negatively charged lipid-containing membrane (28). In light of these results it seems likely that in the presence of lipid II, nisin can translocate across the membrane and thus the inwardly orientated lipid II becomes available for interaction with nisin.…”

Section: Activity Of Wild-type Nisin Z On Lipid Ii-containing Unilamementioning

confidence: 98%

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

Wiedemann¹,

Breukink²,

Kraaij³

et al. 2001

Journal of Biological Chemistry

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667

609

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؊1 for the wildtype 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. [⌬N20/⌬M21]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.The antimicrobial peptide nisin is produced by numerous strains of Lactococcus lactis and inhibits a broad range of Gram-positive bacteria (1, 2). It belongs to the lantibiotics, a group of antimicrobial peptides that is characterized by the presence of intramolecular rings formed by the thioether amino acids lanthionine and 3-methyllanthionine (3, 4). Nisin has had a long history as a potent and safe food preservative, although recent insight into the molecular mechanism of its bactericidal activity also make it interesting for biomedical applications (5, 6). Generally, the nisin-type subgroup of lantibiotics comprises elongated cationic peptides that have the capacity to adopt amphiphilic structures. Such peptides are assumed to kill microbes by disturbing the integrity of the energy-transducing membrane. Indeed, early experiments demonstrated that nisin or related lantibiotics induced rapid efflux of ions or cytoplasmic solutes such as amino acids and nucleotides. The concomitant depolarization of the cytoplasmic membrane resulted in an instant termination of all biosynthetic processes (7,8). Structural analysis in the presence of micelles indicated that the hydrophilic groups of the peptide interact with the phospholipid headgroups, and the hydrophobic side chains are immersed in the hydrophobic core of the membrane (9, 10). The wedge model as proposed by Driessen et al. (11) takes into account such structural data and proposes that the peptides insert into the membrane without losing contact with the membrane surface, resulting in the formation of a short-lived pore.Whereas the wedge model may illustrate results obtained with model membranes, a number of effects observed with intact living cells remain unexplained; in particular, the fact that nisin acts on model membranes at micromolar concentrations whereas in vivo minimal inhibitory concentration (MIC) 1 values are in the nanomolar range. The discrepancies were explained by the finding that nisin and epidermin use lipid II, the bactoprenol-bound precursor of the bacterial cell wall as a docking molecule for subsequent pore formation (12). The specificity of the nisin-lipid II interaction a...

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Section: Discussionsupporting

confidence: 73%

Section: Figmentioning

confidence: 71%

Section: Activity Of Wild-type Nisin Z On Lipid Ii-containing Unilamementioning

confidence: 98%

See 1 more Smart Citation

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

Wiedemann¹,

Breukink²,

Kraaij³

et al. 2001

Journal of Biological Chemistry

Self Cite

667

609

View full text Add to dashboard Cite

show abstract

“…Most type A lantibiotics are cationic, which appears to enhance their interaction with the negatively charged lipid membrane of the target organism. However, introducing more positively charged amino acids into the C-terminus of nisin did not lead to enhanced antimicrobial activity [30,32,33] and might even hinder pore formation [34]. Analogously, we found that adding more C-terminal lysines did not further increase antimicrobial activity of nisin- (1-22).…”

Section: Positive C-terminal Charge Enhances Antimicrobial Activity Omentioning

confidence: 61%

Activity and Export of Engineered Nisin-(1-22) Analogs

Plat¹,

Kuipers²,

Lange³

et al. 2011

Polymers

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Abstract:The pentacyclic peptide antibiotic nisin, produced by Lactococcus lactis is ubiquitously applied as a food preservative. We previously demonstrated that the truncated nisin-(1-22) has only 10-fold lower activity than nisin. Here we aimed at further developing this tricyclic nisin analog to reach activity comparable to that of nisin. Our data demonstrate that: (1) ring A has a large mutational freedom; (2) the composition of residues 20-22 strongly affects production levels of nisin-(1-22); (3) a positively charged C-terminus of nisin-(1-22) significantly enhances its antimicrobial activity; (4) nisin-(1-22) inhibits in vitro growth of a target strain using different dynamics than nisin.

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“…Interestingly, [D]-K 3 H 3 L 9 has no pH-dependent activity, although it contains a significant number of histidines. This differs from several natural antimicrobial peptides that have pH-dependent activity, although they contain only a few histidines in their sequence (36)(37)(38)(39)(40)(41)(42)(43)(44)(45)(46)(47).…”

Section: Discussionmentioning

confidence: 93%

Suppression of Human Solid Tumor Growth in Mice by Intratumor and Systemic Inoculation of Histidine-Rich and pH-Dependent Host Defense–like Lytic Peptides

2009

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Previously, we reported that intratumor or systemic inoculation of a cationic 15-mer, innate immunity-like lytic peptide composed of D-and L-amino acids ([D]-K 6 L 9 ) caused growth arrest of 22RV1 prostate carcinoma xenografts in a mouse model. However, despite its therapeutic potential, this peptide has significant systemic toxicity at concentrations slightly higher than the therapeutic one. Here, we used the acidic environment created by solid tumors as a trigger to activate anticancer lytic peptides by making them cationic only at low pH levels. We achieved this selectivity by substituting lysines (pKa, f10.5) for histidines (pKa, f6.1) in the parental peptide [D]-K 6 L 9 . Histidine is protonated below pH 7. For that purpose, we replaced either three or all six lysines in the parental peptide with histidines to obtain the peptides [D]-K 3 H 3 L 9 and [D]-H 6 L 9 . Interestingly, in vitro experiments showed pH-dependent activity only with [D]-H 6 L 9 mainly toward cancer cell lines. However, both peptides showed reduced systemic toxicity compared with the parental peptide. Intratumor and systemic inoculation of these peptides resulted in a significant decrease in the 22RV1 prostate cancer tumor volume and systemic secretion of prostate-specific antigen in a xenograft mice model. Moreover, histologic modifications revealed a significant reduction in new blood vessels selectively in tumor tissues after treatment with the peptides compared with the untreated tumors. The lytic mode of action of these new peptides, which makes it difficult for the cancer cells to develop resistance, and their selective and pH-dependent activity make them potential candidates for treatment of solid cancer tumors.

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Pore Formation by Nisin Involves Translocation of Its C-Terminal Part across the Membrane

Cited by 77 publications

References 28 publications

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

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

Activity and Export of Engineered Nisin-(1-22) Analogs

Suppression of Human Solid Tumor Growth in Mice by Intratumor and Systemic Inoculation of Histidine-Rich and pH-Dependent Host Defense–like Lytic Peptides

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