Ocellatin‐<scp>PT</scp> antimicrobial peptides: High‐resolution microscopy studies in antileishmania models and interactions with mimetic membrane systems

Oliveira, Margareth Silva; Gomes‐Alves, Ana G.; Sousa, Carla; Marani, Mariela M.; Plácido, Alexandra; Vale, Nuno; Delerue–Matos, Cristina; Gameiro, Paula; Kückelhaus, Selma Aparecida Souza; Tomás, Ana M.; Leite, J. R.; Eaton, Peter

doi:10.1002/bip.22925

Cited by 19 publications

(18 citation statements)

References 40 publications

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“…Leishmania infantum promastigotes were grown at 25°C in RPMI 1640 Glutamax (Gibco), complemented with 10% (v/v) Exosome Depleted FBS (Gibco), 50 U mL-1 penicillin, 50 μ g/mL streptomycin, and 20 mM HEPES sodium salt pH 7.4 (Sigma). After 7–10 days stationary phase cultures Parasites were expanded until they differentiated into infectious metacyclic promastigotes 31 . Parasite culture media was collected at the end of the differentiation, and stored at −20 °C before EV isolation.…”

Section: Methodsmentioning

confidence: 99%

Optimized Isolation of Extracellular Vesicles From Various Organic Sources Using Aqueous Two-Phase System

Kırbaş

Bozkurt

Asutay

et al. 2019

Sci Rep

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From biomarkers to drug carriers, Extracellular Vesicles (EVs) are being used successfully in numerous applications. However, while the subject has been steadily rising in popularity, current methods of isolating EVs are lagging behind, incapable of isolating EVs at a high enough quantity or quality while also requiring expensive, specialized equipment. The “isolation problem” is one of the major obstacles in the field of EV research - and even more so for their potential, widespread use for clinical diagnosis and therapeutic applications. Aqueous Two-Phase Systems (ATPS) has been reported previously as a promising method for isolating EVs quickly and efficiently, and with little contaminants - however, this method has not seen widespread use. In this study, an ATPS-based isolation protocol is used to isolate small EVs from plant, cell culture, and parasite culture sources. Isolated EVs were characterized in surface markers, size, and morphological manner. Additionally, the capacity of ATPS-based EV isolation in removing different contaminants was shown by measuring protein, fatty acid, acid, and phenol red levels of the final isolate. In conclusion, we have shown that EVs originating from different biological sources can be isolated successfully in a cost-effective and user-friendly manner with the use of aqueous two-phase systems.

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Section: Methodsmentioning

confidence: 99%

Optimized Isolation of Extracellular Vesicles From Various Organic Sources Using Aqueous Two-Phase System

Kırbaş

Bozkurt

Asutay

et al. 2019

Sci Rep

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“…The secondary structure of salamandrin-I was assessed using circular dichroism (CD) spectroscopy in the far UV region using a Jasco J-815 CD Spectropolarimeter (Jasco Corp., Tokyo, Japan), as previously reported [20,21]. Briefly, the measurements were carried out under a nitrogen gas flow of 8 L/h at 20 • C. Spectra were obtained between 190 and 260 nm, using a 100 mm cell path length.…”

Section: Circular Dichroismmentioning

confidence: 99%

“…These experiments were performed at 37 • C and a scan speed of 50 nm/min, a response time of 1 s, and a bandwidth of 1 nm were used. The spectra were converted to molar ellipticity per residue, as previously reported [20,21].…”

Section: Circular Dichroismmentioning

confidence: 99%

The Antioxidant Peptide Salamandrin-I: First Bioactive Peptide Identified from Skin Secretion of Salamandra Genus (Salamandra salamandra)

et al. 2020

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Amphibian skin is a multifunctional organ that plays key roles in defense, breathing, and water balance. In this study, skin secretion samples of the fire salamander (Salamandra salamandra) were separated using RP-HPLC and de novo sequenced using MALDI-TOF MS/MS. Next, we used an in silico platform to screen antioxidant molecules in the framework of density functional theory. One of the identified peptides, salamandrin-I, [M + H]+ = 1406.6 Da, was selected for solid-phase synthesis; it showed free radical scavenging activity against DPPH and ABTS radicals. Salamandrin-I did not show antimicrobial activity against Gram-positive and -negative bacteria. In vitro assays using human microglia and red blood cells showed that salamandrin-I has no cytotoxicity up to the concentration of 100 µM. In addition, in vivo toxicity tests on Galleria mellonella larvae resulted in no mortality at 20 and 40 mg/kg. Antioxidant peptides derived from natural sources are increasingly attracting interest. Among several applications, these peptides, such as salamandrin-I, can be used as templates in the design of novel antioxidant molecules that may contribute to devising strategies for more effective control of neurological disease.

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“…Even though there is still structural diversity within each group, the most outstanding source of antileishmanial peptides is the amphibian cluster (30.72%). Skin frog secretions have been described as a therapeutically relevant library of components, including peptides widely capable of inducing the death of micro- and macroscopic pathogens [ 44 , 45 , 46 , 47 ], and with demonstrated activity against both life cycle stages of Leishmania [ 36 , 48 , 49 , 50 ]. Figure 2 B also shows the relevance of structure–function and peptide design studies (7th cluster), as well as the modifications carried out to functionalize peptides for clinically useful applications.…”

Section: Peptides Targeting Leishmania Spp: Current Progressmentioning

confidence: 99%

Peptides to Tackle Leishmaniasis: Current Status and Future Directions

Robles-Loaiza

Pinos-Tamayo

Mendes

et al. 2021

IJMS

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Peptide-based drugs are an attractive class of therapeutic agents, recently recognized by the pharmaceutical industry. These molecules are currently being used in the development of innovative therapies for diverse health conditions, including tropical diseases such as leishmaniasis. Despite its socioeconomic influence on public health, leishmaniasis remains long-neglected and categorized as a poverty-related disease, with limited treatment options. Peptides with antileishmanial effects encountered to date are a structurally heterogeneous group, which can be found in different natural sources—amphibians, reptiles, insects, bacteria, marine organisms, mammals, plants, and others—or inspired by natural toxins or proteins. This review details the biochemical and structural characteristics of over one hundred peptides and their potential use as molecular frameworks for the design of antileishmanial drug leads. Additionally, we detail the main chemical modifications or substitutions of amino acid residues carried out in the peptide sequence, and their implications in the development of antileishmanial candidates for clinical trials. Our bibliographic research highlights that the action of leishmanicidal peptides has been evaluated mainly using in vitro assays, with a special emphasis on the promastigote stage. In light of these findings, and considering the advances in the successful application of peptides in leishmaniasis chemotherapy, possible approaches and future directions are discussed here.

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Ocellatin‐PT antimicrobial peptides: High‐resolution microscopy studies in antileishmania models and interactions with mimetic membrane systems

Cited by 19 publications

References 40 publications

Optimized Isolation of Extracellular Vesicles From Various Organic Sources Using Aqueous Two-Phase System

Optimized Isolation of Extracellular Vesicles From Various Organic Sources Using Aqueous Two-Phase System

The Antioxidant Peptide Salamandrin-I: First Bioactive Peptide Identified from Skin Secretion of Salamandra Genus (Salamandra salamandra)

Peptides to Tackle Leishmaniasis: Current Status and Future Directions

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