Discovery and mechanistic characterization of a structurally-unique membrane active peptide

Bansal, Shivani; Su, Wan-Chih; Budamagunta, Madhu S.; Xiao, Wenwu; Ajena, Yousif; Liu, Ruiwu; Voss, John C.; Carney, Randy P.; Parikh, Atul N.; Lam, Kit S.

doi:10.1016/j.bbamem.2020.183394

Cited by 6 publications

(5 citation statements)

References 50 publications

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“…The formation of tubular lipid-like structures and vesicle budding are typical of several membrane-active peptides such as the recently discovered LBF14 14-mer peptide [ 53 ]. This peptide was found to induce the drastic morphological disruption of membranes, independently of lipid composition.…”

Section: Resultsmentioning

confidence: 99%

“…This peptide was found to induce the drastic morphological disruption of membranes, independently of lipid composition. Its membrane activity was attributed to membrane insertion at the membrane–water interface, inducing local curvatures and membrane remodeling through vesicle bursting and the formation of interconnected tubules and small membrane vesicles [ 53 ].…”

Section: Resultsmentioning

confidence: 99%

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The Unusual Aggregation and Fusion Activity of the Antimicrobial Peptide W-BP100 in Anionic Vesicles

Ferreira

Nunes

et al. 2023

Membranes

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Cationic antimicrobial peptides (CAMPs) offer a promising strategy to counteract bacterial resistance, mostly due to their membrane-targeting activity. W-BP100 is a potent broad-spectrum cecropin-melittin CAMP bearing a single N-terminal Trp, which was previously found to improve its antibacterial activity. W-BP100 has high affinity toward anionic membranes, inducing membrane saturation at low peptide-to-lipid (P/L) ratios and membrane permeabilization, with the unique property of promoting the aggregation of anionic vesicles only at specific P/L ratios. Herein, we aimed to investigate this unusual behavior of W-BP100 by studying its aggregation and fusion properties with negatively-charged large (LUVs) or giant (GUVs) unilamellar vesicles using biophysical tools. Circular dichroism (CD) showed that W-BP100 adopted an α-helical conformation in anionic LUVs, neutralizing its surface charge at the aggregation P/L ratio. Its fusion activity, assessed by Förster resonance energy transfer (FRET) using steady-state fluorescence spectroscopy, occurred mainly at the membrane saturation/aggregation P/L ratio. Confocal microscopy studies confirmed that W-BP100 displays aggregation and detergent-like effects at a critical P/L ratio, above which it induces the formation of new lipid aggregates. Our data suggest that W-BP100 promotes the aggregation and fusion of anionic vesicles at specific P/L ratios, being able to reshape the morphology of GUVs into new lipid structures.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

The Unusual Aggregation and Fusion Activity of the Antimicrobial Peptide W-BP100 in Anionic Vesicles

Ferreira

Nunes

et al. 2023

Membranes

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“…Four OBOC combinatorial linear peptide libraries (8-, 10-, 12-, and 14-mer) were synthesized using 24 building blocks of 16 canonical and 8 non-canonical amino acids according to our previous published method. 16 A subset (∼100 000 beads total) of these libraries were immobilized on a polystyrene plate and sequentially screened for peptide beads binding to fluorescently labeled GUVs of the desired membrane composition. In this case, beads were screened by using a confocal microscope for selective affinity toward fungal membranes over mammalian membranes.…”

Section: ■ Resultsmentioning

confidence: 99%

“…High-performance liquid chromatography was performed on a Waters 2996 HPLC system equipped with a 4.6 × 150 mm Waters Xterra MS C18 5.0 μm column, and it employed a 20 min gradient from 100% aqueous H 2 O (0.1% TFA) to 100% CH 3 CN (0.1% TFA) at a flow rate of 1.0 mL/min. OBOC combinatorial peptide libraries were synthesized according to our previously published methods. ,− PEGA beads (2 g) were washed thoroughly to duly deprotect any existing Fmoc group. Molar excesses (6–7 equiv) of each unique Fmoc-protected amino acid were added to each column of beads, along with a carbodiimide activating agent (Oxyma) and a base (DIC).…”

Section: Methodsmentioning

confidence: 99%

Discovery and Characterization of a Potent Antifungal Peptide through One-Bead, One-Compound Combinatorial Library Screening

Bansal

Liu

et al. 2022

ACS Infect. Dis.

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This work describes the discovery of a bead-bound membrane-active peptide (MAP), LBF127, that selectively binds fungal giant unilamellar vesicles (GUVs) over mammalian GUVs. LBF127 was re-synthesized in solution form and demonstrated to have antifungal activity with limited hemolytic activity and cytotoxicity against mammalian cells. Through systematic structure–activity relationship studies, including N- and C-terminal truncation, alanine-walk, and d-amino acid substitution, an optimized peptide, K-oLBF127, with higher potency, less hemolytic activity, and cytotoxicity emerged. Compared to the parent peptide, K-oLBF127 is shorter by three amino acids and has a lysine at the N-terminus to confer an additional positive charge. K-oLBF127 was found to have improved selectivity toward the fungal membrane over mammalian membranes by 2-fold compared to LBF127. Further characterizations revealed that, while K-oLBF127 exhibits a spectrum of antifungal activity similar to that of the original peptide, it has lower hemolytic activity and cytotoxicity against mammalian cells. Mice infected with Cryptococcus neoformans and treated with K-oLBF127 (16 mg/kg) for 48 h had significantly lower lung fungal burden compared to untreated animals, consistent with K-oLBF127 being active in vivo. Our study demonstrates the success of the one-bead, one-compound high-throughput strategy and sequential screening at identifying MAPs with strong antifungal activities.

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“…In these efforts, a large number of peptides need to be synthesized in small amounts in order to investigate their membrane activity [121,122]. This can be achieved by synthesizing nanomolar amounts of peptides on solid supports [123] following a rational library design approach or in a combinatorial approach, as implemented by split and mix synthesis of picomolar amounts of peptides [124,125]. Split-split methods can be used to systematically shape the sequence space in one bead one compound libraries [126], this way short AMPs were optimized regarding their hemolytic activity.…”

Section: Combinatorial Synthesis Methods To Optimize Membrane Active ...mentioning

confidence: 99%

Design of Membrane Active Peptides Considering Multi-Objective Optimization for Biomedical Application

2022

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A multitude of membrane active peptides exists that divides into subclasses, such as cell penetrating peptides (CPPs) capable to enter eukaryotic cells or antimicrobial peptides (AMPs) able to interact with prokaryotic cell envelops. Peptide membrane interactions arise from unique sequence motifs of the peptides that account for particular physicochemical properties. Membrane active peptides are mainly cationic, often primary or secondary amphipathic, and they interact with membranes depending on the composition of the bilayer lipids. Sequences of these peptides consist of short 5–30 amino acid sections derived from natural proteins or synthetic sources. Membrane active peptides can be designed using computational methods or can be identified in screenings of combinatorial libraries. This review focuses on strategies that were successfully applied to the design and optimization of membrane active peptides with respect to the fact that diverse features of successful peptide candidates are prerequisites for biomedical application. Not only membrane activity but also degradation stability in biological environments, propensity to induce resistances, and advantageous toxicological properties are crucial parameters that have to be considered in attempts to design useful membrane active peptides. Reliable assay systems to access the different biological characteristics of numerous membrane active peptides are essential tools for multi-objective peptide optimization.

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Discovery and mechanistic characterization of a structurally-unique membrane active peptide

Cited by 6 publications

References 50 publications

The Unusual Aggregation and Fusion Activity of the Antimicrobial Peptide W-BP100 in Anionic Vesicles

The Unusual Aggregation and Fusion Activity of the Antimicrobial Peptide W-BP100 in Anionic Vesicles

Discovery and Characterization of a Potent Antifungal Peptide through One-Bead, One-Compound Combinatorial Library Screening

Design of Membrane Active Peptides Considering Multi-Objective Optimization for Biomedical Application

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