Dataset Information

Mass spectrometric identification of antimicrobial peptides from medicinal seeds

ABSTRACT: Traditional medicinal plants contain a variety of bioactive natural products including cysteine-rich (Cys-rich) antimicrobial peptides (AMPs). Cys-rich AMPs are often crosslinked by multiple disulfide bonds which increase their resistance to chemical and enzymatic degradation. However, this class of molecules is relatively underexplored. Herein, in silico analysis predicted 80 – 100 Cys-rich AMPs per species from three edible traditional medicinal plants: Linum usitatissimum (flax), Trifolium pratense (red clover), and Sesamum indicum (sesame). Proteomics analysis of seed peptide extracts revealed direct evidence for the translation of 3-10 Cys-rich AMPs per species including lipid transfer proteins, defensins, α-hairpinins, and snakins. Negative activity revealed by antibacterial screening highlights the importance of employing a multi-pronged approach for AMP discovery. Further, this study demonstrates that flax, red clover, and sesame are promising sources of further AMP discovery and characterization.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Linum Usitatissimum Sesamum Indicum Trifolium Pratense

SUBMITTER: Leslie Hicks

LAB HEAD: Leslie M. Hicks

PROVIDER: PXD029289 | Pride | 2022-02-16

REPOSITORIES: Pride

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Dataset's files

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			Action	DRS
	20210611_AMB_MedPlantSeeds_RAD_LU-80ACN_1x_1uL.mgf	Mgf
	20210611_AMB_MedPlantSeeds_RAD_LU-80ACN_1x_1uL.raw	Raw
	20210611_AMB_MedPlantSeeds_RAD_LU_20ACN_10x-1uL.mgf	Mgf
	20210611_AMB_MedPlantSeeds_RAD_LU_20ACN_10x-1uL.raw	Raw
	20210611_AMB_MedPlantSeeds_RAD_LU_40ACN_10x_1uL.mgf	Mgf

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Publications

Mass Spectrometric Identification of Antimicrobial Peptides from Medicinal Seeds.

Moyer Tessa B TB Brechbill Amanda M AM Hicks Leslie M LM

Molecules (Basel, Switzerland) 20211201 23

Traditional medicinal plants contain a variety of bioactive natural products including cysteine-rich (Cys-rich) antimicrobial peptides (AMPs). Cys-rich AMPs are often crosslinked by multiple disulfide bonds which increase their resistance to chemical and enzymatic degradation. However, this class of molecules is relatively underexplored. Herein, in silico analysis predicted 80-100 Cys-rich AMPs per species from three edible traditional medicinal plants: <i>Linum usitatissimum</i> (flax), <i>Trif ...[more]

PMID: 34885884

Similar Datasets

Project description:Plant cells contain different O-acetylserine(thiol)lyase (OASTL) enzymes involved in Cys biosynthesis and located in different subcellular compartments. These enzymes are made up of a complex variety of isoforms resulting in different subcellular Cys pools. To unravel the contribution of cytosolic Cys to plant metabolism, we characterized the knockout oas-a1.1 and osa-a1.2 mutants, deficient in the most abundant cytosolic OASTL isoform in Arabidposis thaliana. Total intracellular Cys and glutathione concentrations were reduced, and the glutathione redox state was shifted in favour of its oxidized form. Interestingly, the capability of the mutants to chelate heavy metals did not differ from that of the wild type, but the mutants have an enhanced sensitivity to Cd. With the aim of establishing the metabolic network most influenced by the cytosolic Cys pool, we used the ATH1 GeneChip for evaluation of differentially expressed genes in the oas-a1.1 mutant grown under non-stress conditions. The transcriptomic footprints of mutant plants had predicted functions associated with various physiological responses that are dependent on reactive oxygen species and suggested that the mutant was oxidatively stressed. To further elucidate the specific function(s) of the OAS-A1 isoform in the adaptation response to cadmium we extended the trasncriptome experiment to the wild type and oas-a1.1 mutant plants exposed to Cd. The comparison of transcriptomic profiles showed a higher proportion of genes with altered expression in the mutant than in the wild type, highlighting up-regulated genes identified as of the general oxidative stress response rather than metal-responsive genes. Wild type and oas-a1.1 mutant plants were grown hydroponically and, after a two-week acclimation period, the roots and shoots were harvested separately. Total RNA was then prepared and analyzed using the Affymetrix-Arabidopsis ATH1GeneChip array. Three biological replicates were performed for each sample. We made two different comparisons to classify the differently expressed genes in the mutant plant: oas-a1.1 roots versus wild-type roots and oas-a1.1 shoots versus wild-type shoots. Hydroponically-grown wild type and oas-a1.1 mutant plants were further treated with 50µM CdCl2 and 18h-treated-roots and 24h-treated-shoots were harvested. Total RNA was then prepared and analyzed using the Affymetrix-Arabidopsis ATH1GeneChip array. Three biological replicates were performed for each sample. Different comparisons were performed as follows: 18h Cd-treated wild type roots versus untreated wild type roots; 24h Cd-treated wild type shoots versus untreated wild type shoots; 18h Cd-treated oas-a1.1 roots versus untreated oas-a1.1 roots; 24h Cd-treated oas-a1.1 shoots versus untreated oas-a1.1 shoots; 18h Cd-treated oas-a1.1 roots versus 18h Cd-treated wild type roots; 24h Cd-treated oas-a1.1 shoots versus 24h Cd-treated wild type shoots

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.