Cell-penetrating and cargo-delivery ability of a spider toxin-derived peptide in mammalian cells

Ponnappan, Nisha; Chugh, Archana

doi:10.1016/j.ejpb.2017.01.012

Cited by 41 publications

(20 citation statements)

References 36 publications

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“…Spider venom peptides usually affect the permeability of the membrane [32]. LDP-NLS peptide was utilized for the effective delivery of the β-galactosidase enzyme into the HeLa cells [16]. Herein, the LDP-NLS at a molar ratio of 1:10 formed stable nanoparticles through noncovalent binding with the Nef protein.…”

Section: Discussionmentioning

confidence: 99%

“…Preparation and characterization of the Nef protein/LDP complexes. The LDP-NLS amphipathic peptide (KWRRKLKKLRPKKKRKV) [16] was synthesized by Biomatik Corporation (Canada). The formation of non-covalent Nef/LDP-NLS complexes with molar ratios of 1:2, 1:5, 1:10, 1:15, 1:20, and 1:30 (1 μg Nef protein) was performed in PBS and incubated for 60 min at room temperature.…”

Section: Methodsmentioning

confidence: 99%

“…Then, the culture medium was replaced with fresh RPMI 1640. In order to evaluate the cytotoxicity of compounds, the pEGFP-N1-nef (2 μg), Nef protein (1 μg), HR9 (9.3 μg), LDP-NLS (10 μg), and pEGFP-N1-nef/HR9 complexes with an N/P ratio of 5, as well as, Nef/LDP-NLS complexes with a molar ratio of 1:10 [15,16] were added to the cells and incubated for 48 h at 37˚C and 5% CO2 without exchanging the media. After 48 h, the culture medium was removed and the cells were incubated with the MTT (5 mg/ml in PBS) (Sigma-Aldrich, Germany).…”

Section: Methodsmentioning

confidence: 99%

“…Hydrophobic cysteines and histidines help the formation of a pore, and the peptides translocate by diffusing on the surface of the pore [15]. Latarcin 1 peptide (LDP) is an amphipathic peptide derived from Lachesana tarabaevi spider toxin that influences the permeability of biological membranes [16].…”

Section: Vol 7 Nomentioning

confidence: 99%

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In vitro Delivery of HIV-1 Nef Antigen by Histidine-rich nona-arginine and Latarcin 1 peptide

Namazi

Bolhassani

Sadat

et al. 2019

JoMMID

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Introduction: The Nef accessory protein is an attractive antigenic candidate in the development of HIV-1 DNA-or protein-based vaccines. The most crucial disadvantage of DNA and protein-based vaccines is their low immunogenicity, which can be improved by cell-penetrating peptides (CPPs) as effective carrier molecules. Methods: In this study, the HIV-1 Nef protein was generated in the Escherichia coli expression system for in vitro delivery using a novel CPP, Latarcin 1 peptide, in a non-covalent manner. Also, the Histidine-rich nona-arginine peptide was utilized to transfer the HIV-1 Nef gene. The size, morphology, and zeta potential of the complexes were evaluated by scanning electron microscopy (SEM) and Zetasizer. The efficiency of cell transfection was studied using a fluorescence microscopy and flow cytometry for the DNA/CPP complexes and western blot analysis for the protein/CPP complexes. Results: The Nef protein generated in the BL21 strain migrated as a dominant band of ~30 kDa in SDS-PAGE. The SEM data confirmed the formation of stable complexes with a size below 200 nm. MTT assay demonstrated that the complexes did not represent any considerable cytotoxic effect compared to untreated HEK-293T cells. The results of fluorescence microscopy, flow cytometry, and western blotting revealed that the Nef DNA and protein constructs could be significantly transfected into HEK-293T cell line using these CPPs. Conclusion: These data suggest that the Histidine-rich nonaarginine peptide and Latarcin 1 peptide as CPPs can be considered as a promising approach in the development of the HIV-1 vaccine for gene or protein delivery.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Vol 7 Nomentioning

confidence: 99%

See 2 more Smart Citations

In vitro Delivery of HIV-1 Nef Antigen by Histidine-rich nona-arginine and Latarcin 1 peptide

Namazi

Bolhassani

Sadat

et al. 2019

JoMMID

View full text Add to dashboard Cite

show abstract

“…reproducibility and were not suitable for this study. Since the venom was water insoluble, paper discs (as a filter and block) were used in the disc diffusion assay for evaluating the antibacterial activity of the crude venom and its components during culture (28)(29)(30)(31). Some studies have suggested that the antimicrobial effect of venom is promoted by the increased permeability of the bacterial cell membrane induced by antimicrobial proteins present in spider venom (32 (19).…”

Section: Mtt Assay and Mic Determination Disc Diffusion And Well Difmentioning

confidence: 99%

Investigating Antibacterial Effects of Latrodectus Dahli Crude Venom on Escherichia coli, Staphylococcus aureus and Bacillus subtilis

et al. 2019

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Background and Objectives: Nowadays, infections with antibiotic-resistant bacteria are among the most important causes of mortality worldwide. This has attracted the attention of researchers to seek suitable alternatives for antibiotics. The venom of many toxic species such as arthropods has antibacterial properties. In this study, we investigated antibacterial effects of crude venom of Latrodectus dahli on Escherichia coli, Staphylococcus aureus, and Bacillus subtilis.Methods: Lyophilized crude venom of L. dahli was dissolved in 50 mM Tris-HCl buffer. Protein concentration was determined by the Bradford assay. Then, the bacteria were exposed to different concentrations (31.25-250 ng/mL) of the crude venom. Inhibitory activity of the venom against the bacteria was determined by MTT assay and determining minimum inhibitory concentration (MIC).Results: Results of the MTT assay showed that the crude venom significantly inhibited the growth of E. coli (31.25 and 62.5 ng/mL), S. aureus (at 250 ng/mL) and B. subtilis (at 125 and 250 ng/mL). In the MIC experiment, the crude venom significantly inhibited the growth of E. coli (at concentrations of 31.25 and 62.5ng/mL), S. aureus (at concentrations of 31.25-250 ng/mL) and B. subtilis (at concentrations of 31.25-250ng/mL).Conclusion: The crude venom of L. dahli and its components showed relatively strong antibacterial effects.

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A new biology of cell penetrating peptides

Howl

Jones

2020

Peptide Science

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As pharmacokinetic modifiers, cell penetrating peptides (CPPs) have proven utility for the delivery of otherwise impermeable cargoes into the discrete intracellular compartments of eukaryotic cells. Methods for the molecular optimisation of CPP sequences can significantly increase the performance of novel vectors matched to a specific delivery function. Moreover, the ‘information rich’ composition and inherent molecular flexibility of many CPPs can also facilitate their interaction with intracellular proteins and other cellular structures. This proteomimetic property of CPPs has been exploited in the design of bioportides, bioactive CPPs that regulate cellular functions often by binding relatively flat PPI interfaces to achieve a dominant negative action. This new biology of CPPs is rapidly gathering momentum through the design and synthesis of a bewildering variety of peptides that may be, in whole or part, linear, helical, cyclic and/or chimeric in nature. A particular emphasis of contemporary CPP‐centred drug discovery is the unmet medical need of cancer, though the biomedical scope of bioportide applications is impressively broad including cellular signalling and reprogramming. We are hopeful that one or more of the fascinating studies described herein will translate into the clinic to establish bioportide technologies as a viable option for drug discovery.

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Cell-penetrating and cargo-delivery ability of a spider toxin-derived peptide in mammalian cells

Cited by 41 publications

References 36 publications

In vitro Delivery of HIV-1 Nef Antigen by Histidine-rich nona-arginine and Latarcin 1 peptide

In vitro Delivery of HIV-1 Nef Antigen by Histidine-rich nona-arginine and Latarcin 1 peptide

Investigating Antibacterial Effects of Latrodectus Dahli Crude Venom on Escherichia coli, Staphylococcus aureus and Bacillus subtilis

A new biology of cell penetrating peptides

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