35 research outputs found
Recognition of Membrane Sterols by Polyene Antifungals Amphotericin B and Natamycin, A 13C MAS NMR Study
The molecular action of polyene macrolides with antifungal activity, amphotericin B
and natamycin, involves recognition of sterols in membranes. Physicochemical and
functional studies have contributed details to understanding the interactions between
amphotericin B and ergosterol and, to a lesser extent, with cholesterol. Fewer molecular
details are available on interactions between natamycin with sterols. We use solid state
13C MAS NMR to characterize the impact of amphotericin B and natamycin on mixed
lipid membranes of DOPC/cholesterol or DOPC/ergosterol. In cholesterol-containing
membranes, amphotericin B addition resulted in marked increase in both DOPC and
cholesterol 13C MAS NMR linewidth, reflecting membrane insertion and cooperative
perturbation of the bilayer. By contrast, natamycin affects little either DOPC or cholesterol
linewidth but attenuates cholesterol resonance intensity preferentially for sterol core with
lesser impact on the chain. Ergosterol resonances, attenuated by amphotericin B, reveal
specific interactions in the sterol core and chain base. Natamycin addition selectively
augmented ergosterol resonances from sterol core ring one and, at the same time, from
the end of the chain. This puts forward an interaction model similar to the head-to-tail
model for amphotericin B/ergosterol pairing but with docking on opposite sterol faces.
Low toxicity of natamycin is attributed to selective, non-cooperative sterol engagement
compared to cooperative membrane perturbation by amphotericin B
Docking and molecular dynamics simulations of the ternary complex nisin2:lipid II
Lanthionine antibiotics are an important class of naturally-occurring antimicrobial peptides. The best-known, nisin, is a commercial food preservative. However, structural and mechanistic details on nisin/lipid II membrane complexes are currently lacking. Recently, we have developed empirical force-field parameters to model lantibiotics. Docking and molecular dynamics (MD) simulations have been used to study the nisin2:lipid II complex in bacterial membranes, which has been put forward as the building block of nisin/lipid II binary membrane pores. A Ile1Trp mutation of the N-terminus of nisin has been modelled and docked onto lipid II models; the computed binding affinity increased compared to wildtype. Wild-type nisin was also docked onto three different lipid II structures and a stable 2:1 nisin:lipid II complex formed. This complex was inserted into a membrane. Six independent MD simulations revealed key interactions in the complex, specifically the N terminal engagement of nisin with lipid II at the pyrophosphate and C-terminus of the pentapeptide chain. Nisin2 inserts into the membrane and we propose this is the first step in pore formation, mediated by the nisin N-terminus–lipid II pentapeptide hydrogen bond. The lipid II undecaprenyl chain adopted different conformations in the presence of nisin, which may also have implications for pore formation
Passive immunization with Pneumovax®23 and pneumolysin in combination with vancomycin for pneumococcal endophthalmitis
A RNA-seq approach to identify putative toxins from acrorhagi in aggressive and non-aggressive Anthopleura elegantissima polyps
Structural investigations of pneumolysin/lipid complexes and pneumolysin-cholesterol interactions
Interaction of the lantibiotic nisin with mixed lipid bilayers: a 31P and 2H NMR study.
Nisin is a positively charged antibacterial peptide which binds to the negatively charged membranes of Gram-positive bacteria. The initial interaction of the peptide with model membranes of neutral (phosphatidylcholine) and negatively charged (phosphatidylcholine/phosphatidylglycerol) model lipid membranes was studied using nonperturbing solid state magic angle spinning (MAS) (31)P NMR and (2)H wide-line NMR. In the presence of nisin, the coexistence of two bilayer lipid environments was observed both in charged and in neutral membranes. One lipid environment was found to be associated with lipid directly interacting with nisin and one with noninteracting lipid. Solid state (31)P MAS NMR results show that the acidic membrane lipid component partitions preferentially into the nisin-associated environment. Deuterium NMR ((2)H NMR) of the selectively headgroup-labeled acidic lipid provides further evidence of a strong interaction between the charged lipid component and the peptide. The segregation of acidic lipid into the nisin-bound environment was quantified from (2)H NMR measurements of selectively headgroup-deuterated neutral lipid. It is suggested that the observed lipid partitioning in the presence of nisin is driven, at least initially, by electrostatic interactions. (2)H NMR measurements from chain-perdeuterated neutral lipids indicate that nisin perturbs the hydrophobic region of both charged and neutral bilayers
Targeting extracellular pyrophosphates underpins the high selectivity of nisin.
The spread of infectious diseases and the increase in antibiotic resistance represent a life-threatening global development that calls for new approaches to control microorganisms. Of all potential targets, the essential and unique pathway of bacterial cell wall synthesis, targeted by the first known antibiotic penicillin, remains a perfect candidate for the development of new antibiotics. Here we show that the lantibiotic nisin exercises its antibacterial action by targeting peptidoglycan intermediates' extracellular pyrophosphate, unique to bacterial cell wall precursors. We show that nisin sequesters cell wall precursors found in the outer leaflet of bacterial plasma membranes, Lipid II and undecaprenyl pyrophosphate, into stable complexes. We propose a model of antibacterial action for nisin in which the terminal amino group of Ile1 targets the pyrophosphate groups of the bacterial cell wall precursors, where it docks via a hydrogen bond. The pyrophosphate moiety, a highly conserved chemical group different from the L-Lys-D-Ala-D-Ala docking motif for vancomycin, has no biochemical analogs with comparable properties and is unlikely to be susceptible to bacterial adaptations akin to those responsible for resistance to penicillins and vancomycin