Multimerization of a Proline-Rich Antimicrobial Peptide, Chex-Arg20, Alters Its Mechanism of Interaction with the Escherichia coli Membrane

Li, Wenyi; O'Brien-Simpson, Neil M; Tailhades, Julien; Pantarat, Namfon; Dawson, R. M. C.; Ötvös, László; Reynolds, Eric C.; Separovic, Frances; Hossain, Mohammed Akhter; Wade, John D.

doi:10.1016/j.chembiol.2015.08.011

Cited by 59 publications

(90 citation statements)

References 47 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…They kill microorganisms using various strategies that include membrane disruption as the chief mechanism, apart from having downstream intracellular targets (14). They offer themselves as attractive substitutes for conventional small molecule drugs in view of their membrane disruption properties that allow them to circumvent the development of resistance in microbial targets (3,(15)(16)(17)(18). A number of these naturally occurring molecules are endowed with antimicrobial activities; however, their use is restricted to topical applications owing to their cytotoxicity (19).…”

Section: Introductionmentioning

confidence: 99%

Mode of Action of a Designed Antimicrobial Peptide: High Potency against Cryptococcus neoformans

Datta¹,

Yadav

Ghosh³

et al. 2016

Biophysical Journal

View full text Add to dashboard Cite

There is a significant need for developing compounds that kill Cryptococcus neoformans, the fungal pathogen that causes meningoencephalitis in immunocompromised individuals. Here, we report the mode of action of a designed antifungal peptide, VG16KRKP (VARGWKRKCPLFGKGG) against C. neoformans. It is shown that VG16KRKP kills fungal cells mainly through membrane compromise leading to efflux of ions and cell metabolites. Intracellular localization, inhibition of in vitro transcription, and DNA binding suggest a secondary mode of action for the peptide, hinting at possible intracellular targets. Atomistic structure of the peptide determined by NMR experiments on live C. neoformans cells reveals an amphipathic arrangement stabilized by hydrophobic interactions among A2, W5, and F12, a conventional folding pattern also known to play a major role in peptide-mediated Gram-negative bacterial killing, revealing the importance of this motif. These structural details in the context of live cell provide valuable insights into the design of potent peptides for effective treatment of human and plant fungal infections.

show abstract

Section: Introductionmentioning

confidence: 99%

Mode of Action of a Designed Antimicrobial Peptide: High Potency against Cryptococcus neoformans

Datta¹,

Yadav

Ghosh³

et al. 2016

Biophysical Journal

View full text Add to dashboard Cite

show abstract

“…In vitro cytotoxicity was also measured via the Promega CellTiter 96 AqueousNon-Radioactive Cell Proliferation Assay (Li et al, 2015a) using the mammalian cell lines HEK-293 (ATCC CRL 1573) and H-4-II-E (ATCC CRL-1548). None of the Chex1-Arg20 analogs showed any toxicity against either mammalian cell line at the highest tested concentration (100 μM) (Table 3).…”

Section: Resultsmentioning

confidence: 99%

“…The peptides were synthesized by Fmoc/tBu solid-phase methods (Fields and Noble, 1990) using a CEM Liberty microwave-assisted synthesizer and TentaGel-MB-RAM-resin as previously described (Li et al, 2015a). Standard Fmoc-chemistry was used throughout with a 4-fold molar excess of the Fmoc-protected amino acids in the presence of 4-fold HCTU and 8-fold DIPEA.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

The Effect of Selective D- or Nα-Methyl Arginine Substitution on the Activity of the Proline-Rich Antimicrobial Peptide, Chex1-Arg20

Sun

O'Brien-Simpson

et al. 2017

Front. Chem.

Self Cite

View full text Add to dashboard Cite

In vivo pharmacokinetics studies have shown that the proline-rich antimicrobial peptide, A3-APO, which is a discontinuous dimer of the peptide, Chex1-Arg20, undergoes degradation to small fragments at positions Pro6-Arg7 and Val19-Arg20. With the aim of minimizing or abolishing this degradation, a series of Chex1-Arg20 analogs were prepared via Fmoc/tBu solid phase peptide synthesis with D-arginine or, in some cases, peptide backbone Nα-methylated arginine, substitution at these sites. All the peptides were tested for antibacterial activity against the Gram-negative bacterium Klebsiella pneumoniae. The resulting activity of position-7 substitution of Chex1-Arg20 analogs showed that arginine-7 is a crucial residue for maintaining activity against K. pneumoniae. However, arginine-20 substitution had a much less deleterious effect on the antibacterial activity of the peptide. Moreover, none of these peptides displayed any cytotoxicity to HEK and H-4-II-E mammalian cells. These results will aid the development of more effective and stable PrAMPs via judicious amino acid substitutions.

show abstract

“…This information may then be used to retrieve information on the molecular mode of action, target, and specificity of the AMPs [157,[254][255][256]. In addition to addressing the mechanism of cell entry, FC is an especially reliable and robust method in the investigation of CPP internalization and subsequent intracellular trafficking to quantify the fraction of internalized CPPs [159].…”

Section: Live Imagingmentioning

confidence: 99%

Membrane Active Peptides and Their Biophysical Characterization

Avcı

Akbulut

Özkırımlı

2018

Preprint

View full text Add to dashboard Cite

In the last 20 years, an increasing number of studies have been reported on membrane active peptides, which exert their biological activity by interacting with the cell membrane either to disrupt it and lead to cell lysis or to translocate through it to deliver cargos into the cell and reach their target. These peptides are attractive alternatives to currently used pharmaceuticals. Antimicrobial peptides (AMPs) and peptides designed for drug and gene delivery currently in the drug pipeline suggest that these membrane active peptides will soon constitute a significant percentage of the drug market. Here, we focus on two most prominent classes of membrane active peptides; AMPs and cell-penetrating peptides (CPPs). AMPs are a group of membrane active peptides that disrupt the membrane integrity or inhibit the cellular functions of bacteria, virus and fungi. CPPs are another group of membrane active peptides that mainly function as cargo-carriers even though they may also show antimicrobial activity to some extent. Biophysical techniques to understand how they interact with the membrane have shed light on the peptide-membrane interaction at various levels of detail. Structural investigation of membrane active peptides in the presence of the membrane provides important clues on the effect of the membrane environment on peptide conformations. Advances in live imaging techniques have allowed examination of peptide action at a single cell or single molecule level. In addition to these experimental biophysical techniques, molecular dynamics simulations provided clues on the peptide-lipid interactions and dynamics of the cell entry process at atomic detail. In this review, we summarize the recent advances in experimental and computational investigation of membrane active peptides with particular emphasis on two amphipathic membrane active peptides, the AMP melittin and the CPP pVEC.

show abstract

Multimerization of a Proline-Rich Antimicrobial Peptide, Chex-Arg20, Alters Its Mechanism of Interaction with the Escherichia coli Membrane

Cited by 59 publications

References 47 publications

Mode of Action of a Designed Antimicrobial Peptide: High Potency against Cryptococcus neoformans

Mode of Action of a Designed Antimicrobial Peptide: High Potency against Cryptococcus neoformans

The Effect of Selective D- or Nα-Methyl Arginine Substitution on the Activity of the Proline-Rich Antimicrobial Peptide, Chex1-Arg20

Membrane Active Peptides and Their Biophysical Characterization

Contact Info

Product

Resources

About