Role of membrane sterol and redox system in the anti-candida activity reported for Mo-CBP2, a protein from Moringa oleifera seeds

Neto, João Xavier da Silva; Costa, Helen Paula Silva da; Vasconcelos, Ilka M.; Pereira, Mirella Leite; Oliveira, José T.A.; Lopes, Tiago Deiveson Pereira; Dias, Lucas Pinheiro; Araújo, Nadine Monteiro Salgueiro; Moura, Luiz Francisco Wemmenson Gonçalves; Tilburg, Maurício Fraga van; Guedes, Maria Izabel Florindo; Lopes, Larissa Alves; Morais, Eva Gomes; Sousa, Daniele de Oliveira Bezerra de

doi:10.1016/j.ijbiomac.2019.09.142

Cited by 15 publications

(13 citation statements)

References 85 publications

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“…The most affected peptide, RcAlb-PepII, completely lost its activity. Similar results were posted by Neto et al [20] for an anticandidal protein that had its activity reduced by 60% in the presence of AsA. Fluorescence microscopy (Fig.…”

Section: Discussionsupporting

confidence: 87%

“…There are not many studies with similar results to ours to redox enzymes with peptides on yeasts, even to C. neoformans. However, Neto et al [20] reported that a chitinbinding protein purified from Moringa oleifera seeds also caused perturbation in the redox enzymes leading to the accumulation of ROS. In that case, the authors only measured the accumulation of H2O2.…”

Section: Discussionmentioning

confidence: 99%

“…In addition, the same assay to detect H2O2 was performed in the presence of 10 mM ascorbic acid (AsA) [20]. Besides, the pore formation in the presence and absence of 10 mM AsA using the Propidium Iodide (PI) influx assay, following the methodology described in [7].…”

Section: Detection Of Ros Overproductionmentioning

confidence: 99%

“…The assay was performed according to [33] to evaluate the catalase activity. After the antifungal assay at the same conditions described previously, cells were washed three times with 0.15 M NaCl, resuspended in 0.05 M sodium acetate buffer pH 5.2, frozen for 24 h, sonicated for 30 min, centrifuged for 10 min, 10000 g at 4 °C and the supernatant collected as described by [20]. 200 μL of samples were incubated with 700 μL phosphate buffer of 50 mM potassium, pH 7.0 at 30 °C for 10 min.…”

Section: Catalase (Cat)mentioning

confidence: 99%

“…The ergosterol biosynthesis inhibition was evaluated following the method described previously by Neto et al [20]. The cryptococcal cells were cultured in YPD medium in the presence of synthetic peptides (MIC50), 5% DMSO, and Itraconazole (1000 µg mL -1 ) for 24 h, 30 °C.…”

Section: Ergosterol Biosynthesis Inhibitionmentioning

confidence: 99%

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Behind the Curtain: In silico and in vitro Experiments Brought to Light New Insights Into the Anticryptococcal Action of Synthetic Peptides

Aguiar¹,

Neto²,

Silva³

et al. 2022

Preprint

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Cryptococcus neoformans threaten the health, causing cryptococcal meningitis and pneumonia, especially in immunosuppressed patients, which can be fatal. Recently, our research group evaluated and studied the mechanisms of action of four synthetic peptides (SP) against C. neoformans. Here, in silico and in vitro analyses help deepen understanding of peptides' mechanisms of action. The interaction of the peptides with a membrane receptor was analyzed by docking analysis, in addition to ROS overproduction and the modulation of redox metabolism, inhibition of ergosterol biosynthesis, and release of cytochrome c. Out of four, three peptides interacted with membrane receptor PHO36 altering its structure and function and leading to a higher accumulation of O₂- and H2O2. C. neoformans cells treated with SP presented a reduction in the activity of antioxidant enzymes, corroborating ROS accumulation. However, in the presence of the antioxidant ascorbic acid, some peptides could not induce this oxidative stress and have the activity against C. neoformans affected. Curiously, two of these SPs still maintained the activity against C. neoformans and even induced the membrane pore formation as revealed by propidium iodide uptake assay, revealing their mechanism of action is ROS-independent. Additionally, SPs inhibited the biosynthesis of ergosterol, which corroborates the pore formation on the membrane of C. neoformans cells, inhibited the lactate dehydrogenase activity affecting the cell metabolism, and induced the release of Cyt c from the mitochondria inducing death by apoptosis in the cryptococcal cells. Our findings strongly suggest that SPs act by multiple mechanisms, making it difficult for C. neoformans to acquire resistance highlighting the potential of SPs as alternative molecules in treating infections caused by C. neoformans.

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Section: Discussionsupporting

confidence: 87%

Section: Discussionmentioning

confidence: 99%

Section: Detection Of Ros Overproductionmentioning

confidence: 99%

Section: Catalase (Cat)mentioning

confidence: 99%

Section: Ergosterol Biosynthesis Inhibitionmentioning

confidence: 99%

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Behind the Curtain: In silico and in vitro Experiments Brought to Light New Insights Into the Anticryptococcal Action of Synthetic Peptides

Aguiar¹,

Neto²,

Silva³

et al. 2022

Preprint

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Chitin‐binding peptides from Capsicum annuum with antifungal activity and low toxicity to mammalian cells and Galleria mellonella larvae

Gonçalves,

Silva,

dos Santos

et al. 2024

Peptide Science

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In recent years, there have been several reports of the presence of toxic proteins in cultivated or wild plant species, which are implicated in plant defense mechanisms. The existence of these proteins raises the possibility of biotechnological applications originating from the development of new techniques to combat diseases caused by fungi. In this context, there are chitin‐binding proteins. Chitin is an essential component of the fungal cell wall, so chitin‐binding proteins are important in controlling fungal growth. Thus, the objective of this study was to characterize and evaluate the in vitro antimicrobial effect of peptides with chitin binding properties isolated from Capsicum annuum seeds on the growth of the genus Candida. Initially, proteins were extracted in phosphate pH 5.4, and a chitin column was equilibrated with sodium acetate (0.08 M, pH 4.5), where 50 mg of the peptide‐rich heated fraction from each species was applied. Subsequently, the retained material was eluted with 0.1 M HCl. Tricine SDS–PAGE was used to visualize the peptides. After chromatography, two fractions, F1 (not retained in the chitin column) and F2 (retained in the chitin column, named Ca‐F2), were obtained. Electrophoresis showed major protein bands between 3 and 14 kDa. Electrophoresis from chitin affinity chromatography also showed major bands between 3 and 14 kDa, especially for Ca‐F2 retained in the column. One peptide obtained from the F2 fraction was identified by mass spectrometry and showed similarity to seed 2S albumin, named Ca‐Alb2S. Ca‐F2 inhibited the growth of C. albicans and C. tropicalis, was not toxic to mammalian cells and still had a high survival rate when tested in vivo on Galleria mellonella larvae. This is the first report of chitin‐binding peptides isolated from Capsicum seeds through an affinity column and their biological activities. These studies are at an early stage; therefore, other tests are needed to study the mechanism of action of the fraction, since the findings indicate great potential for the development of new antifungal molecules.

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Computer-made peptide RQ18 acts as a dual antifungal and antibiofilm peptide though membrane-associated mechanisms of action

Almeida,

de Oliveira,

Almeida

et al. 2024

Archives of Biochemistry and Biophysics

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Role of membrane sterol and redox system in the anti-candida activity reported for Mo-CBP2, a protein from Moringa oleifera seeds

Cited by 15 publications

References 85 publications

Behind the Curtain: In silico and in vitro Experiments Brought to Light New Insights Into the Anticryptococcal Action of Synthetic Peptides

Behind the Curtain: In silico and in vitro Experiments Brought to Light New Insights Into the Anticryptococcal Action of Synthetic Peptides

Chitin‐binding peptides from Capsicum annuum with antifungal activity and low toxicity to mammalian cells and Galleria mellonella larvae

Computer-made peptide RQ18 acts as a dual antifungal and antibiofilm peptide though membrane-associated mechanisms of action

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