Dataset Information

MoCBP3, an Antifungal Chitin-Binding Protein from Moringa oleifera Seeds, is a Member of the 2S Albumin Family

ABSTRACT: MoCBP3 is a chitin-binding protein from M. oleifera seeds that inhibits the germination and mycelial growth of phytopathogenic fungi. This protein is highly thermostable and resistant to pH changes, and therefore may be useful in the development of new antifungal drugs. However, the relationship of MoCBP3 with the known families of carbohydrate-binding domains has not been established. In the present study, full-length cDNAs encoding 4 isoforms of MoCBP3 (MoCBP3-1, MoCBP3-2, MoCBP3-3 and MoCBP3-4) were cloned from developing seeds. The polypeptides encoded by the MoCBP3 cDNAs were predicted to contain 160 (MoCBP3-3) and 163 amino acid residues (MoCBP3-1, MoCBP3-2 and MoCBP3-4) with a signal peptide of 20-residues at the N-terminal region. A comparative analysis of the deduced amino acid sequences revealed that MoCBP3 is a typical member of the 2S albumin family, as shown by the presence of an eight-cysteine motif, which is a characteristic feature of the prolamin superfamily. Furthermore, mass spectrometry analysis demonstrated that MoCBP3 is a mixture of isoforms that correspond to different mRNA products. The identification of MoCBP3 as a genuine member of the 2S albumin family reinforces the hypothesis that these seed storage proteins are involved in plant defense. Moreover, the chitin-binding ability of MoCBP3 reveals a novel functionality for a typical 2S albumin.

INSTRUMENT(S): Synapt MS

ORGANISM(S): Moringa Oleifera (horseradish Tree) (moringa Pterygosperma)

TISSUE(S): Plant Cell, Cotyledon

SUBMITTER: FREDERICO MORENO

LAB HEAD: Thalles Barbosa Grangeiro

PROVIDER: PXD001762 | Pride | 2016-05-25

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

Items per page:

1 - 5 of 10

Publications

Mo-CBP3, an antifungal chitin-binding protein from Moringa oleifera seeds, is a member of the 2S albumin family.

Freire José E C JE Vasconcelos Ilka M IM Moreno Frederico B M B FB Batista Adelina B AB Lobo Marina D P MD Pereira Mirella L ML Lima João P M S JP Almeida Ricardo V M RV Sousa Antônio J S AJ Monteiro-Moreira Ana C O AC Oliveira José T A JT Grangeiro Thalles B TB

PloS one 20150319 3

Mo-CBP3 is a chitin-binding protein from M. oleifera seeds that inhibits the germination and mycelial growth of phytopathogenic fungi. This protein is highly thermostable and resistant to pH changes, and therefore may be useful in the development of new antifungal drugs. However, the relationship of MoCBP3 with the known families of carbohydrate-binding domains has not been established. In the present study, full-length cDNAs encoding 4 isoforms of Mo-CBP3 (Mo-CBP3-1, Mo-CBP3-2, Mo-CBP3-3 and Mo ...[more]

PMID: 25789746

Similar Datasets

Project description:Antimicrobial peptides (AMPs) are a group of molecules that occur naturally in all living organisms and help them to defend against various pathogens, including fungi, bacteria and viruses (Gan et al., 2021). AMPs have a low molecular mass (~2-50 amino acids), are rich in hydrophobic amino acids and have a net positive charge (Huan et al., 2020). Some stable antimicrobial peptides can bind to chitin, an insoluble homopolymer present in several types of biological materials, such as the cell wall of fungi, insects and crustacean shells; however, chitin is not present in plants. Some peptides, including chitinases, vicilins, lectins, 2S albumins and hevein-like peptides, can bind to a matrix composed of chitin. These peptides exhibit antimicrobial activity against bacteria and fungal pathogens, and they interact with fungal chitin through the chitin binding site, which plays a significant role in inhibiting fungal growth (Slezina and Odintsova, 2023). The binding site shares structural similarity with hevein, an antimicrobial peptide from the latex of Hevea brasiliensis (Willd. exA. Juss.) Müll. Arg. The main element of all hevein-like peptides is that a conservative chitin-binding site is one of their essential structural modules (Slavokhotova et al., 2017; Azmil et al., 2021). Hevein-like peptides are AMPs that are rich in cysteine, contain 29-45 amino acids and 5-7 residues contain glycine. A main feature of hevein-like peptides is a conservative chitin binding site with the amino acid sequence SXFGY/SXYGY, in which X is any amino acid residue (Beintema, 1994). In addition, these peptides are present in the plant cysteine-rich antimicrobial peptide superfamily, are 2-6 kDa in weight, contain 6-10 Cys residues and contain three to five disulfide bridges (Tam et al., 2015). They also possess structural motifs composed of three antiparallel beta sheets and a short α-helix, which are stabilized by 3-5 disulfide bonds (Odintsova et al., 2020). Previously, hevein-like peptides have been reported to exhibit activity against several fungal microorganisms with or without chitin in their cell wall (Slezina and Odintsova, 2023). The mechanism underlying the activity of hevein-like peptides is mainly through depolarization of hyphae membrane (Azmil et al., 2021). Recently, several hevein-like peptides have been isolated from several plant species, such as wheat (Odintsova et al., 2020), quinoa (Loo et al., 2021) and moringa (Sousa et al., 2020). These peptides exhibited activity against a range of microorganisms, such as F. oxysporum, F. culmorum, Bipolaris sorokininana, Alternaria alternata and Cladosporium cucumerinum (Odintsova et al., 2020). These findings have encouraged further studies on the application of these molecules against pathogenic fungi, as the molecules exhibit great activity and could be used to create new antifungal medications. Peppers and chilies originate from the tropical Americas and belong to 22 families of Solanaceae and the genus Capsicum; 35 of these species have been domesticated, while the others remain semidomesticated and wild. The plants are dispersed worldwide, and in Brazil, they are cultivated mainly in the states of Rio de Janeiro, Minas Gerais, Bahia and Goiás; among the various locations, Rio de Janeiro is a special center of diversity (Moscone et al., 2006). These plants are essential solanaceous vegetables with high world economic value. However, the growth, quality and yield of these plants are reduced by biotic (fungal, bactéria and virus infections and pests) and abiotic (drought, salinity and extreme temperatures) factors (Mittler, 2006; Wang et al., 2003). Several AMPs have been isolated from different organs of plants within the Capsicum genus (Oliveira et al., 2022). For example, a hevein-like peptide was identified from C. annuum leaves; this peptide is approximately 4.2 kDa and was named HEVCAN. In addition, the peptide belongs to the chitin-binding domain family and exhibits antimicrobial activity (Oliveira et al., 2022). Our research group has previously isolated and identified several other antimicrobial peptides, such as defensin (Gebara et al., 2020), LTP (Diz et al., 2011), protease inhibitors (Ribeiro et al., 2013) and chitin-binding peptides (Gonçalves et al., 2024), from Capsicum seeds. In this study, we used a chitin affinity column to purify peptide fractions from C. chinense seeds, and these fractions exhibited chitin-binding capacity and activity against yeasts.

Project description:Purpose: Evaluate the anti-tumorigenic efficacy and immunomodulatory effects of chitin as general blocker of immunosuppressive chitinase-like proteins (CLPs) in combination with and without anti-PD-1 immune checkpoint blockade (ICB) in an immunocompetent 4T1- and 66cl4-based intraductal model for triple-negative breast cancer (TNBC). Methods: 4T1 and 66cl4 mammary tumor-bearing female BALB/c mice were either left untreated or were treated with anti-PD-1, chitin or chitin + anti-PD-1 for 2 weeks (w) between 3 and 5 w post-inoculation (p.i.) of the mammary tumor cells. After the 2-w treatment, 4T1 and 66cl4 primary tumors were resected and RNA was isolated from the tissue using in-house developed protocols. Results: Chitin-mediated reduction in innate immunosuppression and increased anti-tumor T-cell immunity in primary tumors of both TNBC models was clearly presented at the genomic level based on RNA-sequencing analysis. More specifically, a selection of 68 and 55 innate immunity-related genes in respectively 4T1 and 66cl4 primary tumors were downregulated upon chitin treatment in combination with or without anti-PD-1. Several of the selected innate immunity genes could be linked to immunosuppressive myeloid cell types, including Ccl2, Cxcl2, Csf1r and Itgam. Genes related to T-cell exhaustion, including Pdcd1, Cd274, Havcr2 and Lag3, were downregulated upon chitin and chitin + anti-PD-1 treatment in 4T1 tumors. Chitin- and chitin + anti-PD-1-treated 66cl4 tumors showed upregulation of genes associated with enhanced T-cell activity, including Cd8a, Cd8b1, Ifng, Il12b and Il18r1. In line with an overall reduced inflammation upon chitin treatment at the protein level, predominantly through myeloid cell reduction, a selected set of 68 and 48 inflammation-related genes were downregulated in chitin- and chitin + anti-PD-1-treated primary tumors of respectively the 4T1- and the 66cl4-based model. Importantly, genes that have been associated with the signaling and pro-tumorigenic activity of a prominent CLP family member, i.e. CHI3L1, were also downregulated in primary tumors following chitin treatment either with or without anti-PD-1, including Usf1 and Rab37 in the 4T1-based model and Lgals3 in the 66cl4-based model. Conclusion: Our findings highlight that chitin, as a general CLP blocker, reduces cancer-associated immunosuppression and enhances anti-tumor immunity as well as ICB responses in complementary ICB-resistant TNBC models, supporting its potential clinical relevance in immunosuppressed TNBC patients.

			Action	DRS
	4KDa.pkl	Other
	4KDa.pride.mgf.gz	Mgf
	4KDa.zip	Other
	8KDa.pkl	Other
	8KDa.pride.mgf.gz	Mgf