Electrophysiological Analysis of Membrane Disruption by Bombinin and Its Isomer Using the Lipid Bilayer System

Sekiya, Yusuke; Shimizu, Keisuke; Kitahashi, Yuki; ohyama, akifumi; Kawamura, Izuru; Kawano, Ryuji

doi:10.1021/acsabm.8b00835

Cited by 24 publications

(30 citation statements)

References 32 publications

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“…Charge flux (CF) through the nanopore reflected the ion flux (Figure 4c) and exhibited similar behavior to that of lipid bilayer leakage. 32 Therefore, it is strongly correlated with membrane dysfunction-related antimicrobial activity. In these measurements, we set the threshold value at 1000 pA; this value was established in reference to our previous study of CF measurements.…”

Section: Resultsmentioning

confidence: 99%

Electrophysiological Analysis of Antimicrobial Peptides in Diverse Species

2019

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This study describes a technical platform that allows us to measure the pore-forming activity of antimicrobial peptides (AMPs) in the lipid bilayer and estimate antimicrobial activity. We selected six different AMPs of diverse species from urochordata to vertebrata and measured the channel current signals using a microfabricated lipid bilayer system. As a result of the electrophysiological measurements, we were able to estimate the pore-forming activity and roughly predict the antimicrobial activity although there was not a strong correlation between the pore-forming activity and the variety of species. Our method will be a unique tool for analyzing a wide variety of diverse AMPs.

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Section: Resultsmentioning

confidence: 99%

Electrophysiological Analysis of Antimicrobial Peptides in Diverse Species

2019

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“…The electrophysiological measurement of AMPs could be a useful method for observing and analyzing pore formation using the droplet contact method ( Figure 3 a,b) [ 16 , 17 ]. The representative observations of electrophysiological measurements are shown in Figure 3 c. Briefly, cell penetrating peptides (CPPs) exhibit step-like signals [ 16 ].…”

Section: Resultsmentioning

confidence: 99%

“…The representative observations of electrophysiological measurements are shown in Figure 3 c. Briefly, cell penetrating peptides (CPPs) exhibit step-like signals [ 16 ]. In contrast, the toroidal pore model exhibits multilevel current signals [ 17 , 18 ]. Based on this knowledge, we performed electrophysiological measurement against 1,2-dioleoyl- sn -glycero-3-phosphoethanolamine (DOPE)/1,2-dioleoyl- sn -glycero-3-phospho-(1′- rac -glycerol) (DOPG) and 1,2-dioleoyl- sn -glycero-3-phosphocholine (DOPC) bilayer membranes, which mimic the microbial and mammalian cell membranes, respectively, to investigate the action mechanisms on the antimicrobial and hemolytic activities of peptides 1 , 2 , and 8 .…”

Section: Resultsmentioning

confidence: 99%

Development of Antimicrobial Stapled Peptides Based on Magainin 2 Sequence

et al. 2021

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Magainin 2 (Mag2), which was isolated from the skin of the African clawed frog, is a representative antimicrobial peptide (AMP) that exerts antimicrobial activity via microbial membrane disruption. It has been reported that the helicity and amphipathicity of Mag2 play important roles in its antimicrobial activity. We investigated and recently reported that 17 amino acid residues of Mag2 are required for its antimicrobial activity, and accordingly developed antimicrobial foldamers containing α,α-disubstituted amino acid residues. In this study, we further designed and synthesized a set of Mag2 derivatives bearing the hydrocarbon stapling side chain for helix stabilization. The preferred secondary structures, antimicrobial activities, and cell-membrane disruption activities of the synthesized peptides were evaluated. Our analyses revealed that hydrocarbon stapling strongly stabilized the helical structure of the peptides and enhanced their antimicrobial activity. Moreover, peptide 2 stapling between the first and fifth position from the N-terminus showed higher antimicrobial activity than that of Mag2 against both gram-positive and gram-negative bacteria without exerting significant hemolytic activity. To investigate the modes of action of tested peptides 2 and 8 in antimicrobial and hemolytic activity, electrophysiological measurements were performed.

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“…We have previously proposed current signal classification for several different signal shapes, and assigned these to various pore-forming models of α-helical peptides. [50][51][52] In this study, we also classified these signals into four types of current signals: step, square-top, multi-level, and erratic. The definition of signal classification is described in Fig.…”

Section: Confirmation Of Pore Formation Using Channel Current Measurementioning

confidence: 99%

De Novo Design of a Nanopore for DNA Detection Incorporating a β-hairpin Peptide

Shimizu¹,

Mijiddorj²,

Yoshida³

et al. 2020

Preprint

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The amino acid sequence of a protein encodes information on its three-dimensional structure and specific functionality. De novo protein design has emerged as a method to manipulate the primary structure for the development of artificial proteins and peptides with desired functionality. This paper describes the de novo design of a pore-forming peptide that has a β-hairpin structure and assembles to form a stable nanopore in a bilayer lipid membrane. This large synthetic nanopore is an entirely artificial device with practical applications. This peptide, named SV28, forms nanopore structures ranging from 1.6 to 6.2 nm in diameter assembled from 7 to 18 monomers. The nanopore formed with a diameter of 5 nm is able to detect long double-stranded DNA (dsDNA) with 1 kbp length, and measurement of current signals allowed us to investigate the translocation behavior of dsDNA at the single molecule level. Such de novo design of peptide sequences has the potential to create assembled structure in lipid membrane such as novel nanopores, which would also be applicable in molecular transporter between inside and outside of lipid membrane.

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Electrophysiological Analysis of Membrane Disruption by Bombinin and Its Isomer Using the Lipid Bilayer System

Cited by 24 publications

References 32 publications

Electrophysiological Analysis of Antimicrobial Peptides in Diverse Species

Electrophysiological Analysis of Antimicrobial Peptides in Diverse Species

Development of Antimicrobial Stapled Peptides Based on Magainin 2 Sequence

De Novo Design of a Nanopore for DNA Detection Incorporating a β-hairpin Peptide

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