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1.
Funct Integr Genomics ; 24(5): 142, 2024 Aug 27.
Artículo en Inglés | MEDLINE | ID: mdl-39187716

RESUMEN

Parsley is a commonly cultivated Apiaceae species of culinary and medicinal importance. Parsley has several recognized health benefits and the species has been utilized in traditional medicine since ancient times. Although parsley is among the most commonly cultivated members of Apiaceae, no systematic genomic research has been conducted on parsley. In the present work, parsley genome was sequenced using the long-read HiFi (high fidelity) sequencing technology and a draft contig assembly of 1.57 Gb that represents 80.9% of the estimated genome size was produced. The assembly was highly repeat-rich with a repetitive DNA content of 81%. The assembly was phased into a primary and alternate assembly in order to minimize redundant contigs. Scaffolds were constructed with the primary assembly contigs, which were used for the identification of AMP (antimicrobial peptide) genes. Characteristic AMP domains and 3D structures were used to detect and verify antimicrobial peptides. As a result, 23 genes (PcAMP1-23) representing defensin, snakin, thionin, lipid transfer protein and vicilin-like AMP classes were identified. Bioinformatic analyses for the characterization of peptide physicochemical properties indicated that parsley AMPs are extracellular peptides, therefore, plausibly exert their antimicrobial effects through the most commonly described AMP action mechanism of membrane attack. AMPs are attracting increasing attention since they display their fast antimicrobial effects in small doses on both plant and animal pathogens with a significantly reduced risk of resistance development. Therefore, identification and characterization of AMPs is important for their incorporation into plant disease management protocols as well as medicinal research for the treatment of multi-drug resistant infections.


Asunto(s)
Petroselinum , Petroselinum/genética , Péptidos Antimicrobianos/genética , Péptidos Antimicrobianos/farmacología , Péptidos Antimicrobianos/metabolismo , Péptidos Antimicrobianos/química , Secuenciación Completa del Genoma , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Genoma de Planta
2.
J Sci Food Agric ; 104(4): 2467-2476, 2024 Mar 15.
Artículo en Inglés | MEDLINE | ID: mdl-37986244

RESUMEN

BACKGROUND: The application of curcumin (Cur) in the food industry is usually limited by its low water solubility and poor stability. This study aimed to fabricate self-assembled nanoparticles using pea vicilin (7S) through a pH-shifting method (pH 7-pH 12-pH 7) to develop water-soluble nanocarriers of Cur. RESULTS: Intrinsic fluorescence, far-UV circular dichroism spectra and transmission electron microscopy analysis demonstrated that the structure of 7S could be unfolded at pH 12.0 and refolded when the pH shifted to 7.0. The assembled 7S-Cur exhibited a high loading ability of 81.63 µg mg-1 for Cur and homogeneous particle distribution. Cur was encapsulated in the 7S hydrophobic nucleus in an amorphous form and combined through hydrophobic interactions and hydrogen bonding, resulting in the static fluorescence quenching of 7S. Compared with free Cur, the retention rates of Cur in 7S-Cur were approximately 1.12 and 1.70 times higher under UV exposure at 365 nm or heating at 75 °C for 120 min, respectively, as well as 7S-Cur showing approximately 1.50 times higher antioxidant activity. During simulated gastrointestinal experiments, 7S-Cur exhibited a better sustained-release property than free Cur. CONCLUSION: The self-assembled 7S nanocarriers prepared using a pH-shifting method effectively improved the antioxidant activity, environmental stability and sustained-release property of Cur. Therefore, 7S isolated from pea protein could be used as potential nanocarriers for Cur. © 2023 Society of Chemical Industry.


Asunto(s)
Curcumina , Nanopartículas , Proteínas de Almacenamiento de Semillas , Curcumina/química , Antioxidantes , Pisum sativum , Preparaciones de Acción Retardada , Portadores de Fármacos/química , Nanopartículas/química , Agua , Tamaño de la Partícula
3.
Crit Rev Food Sci Nutr ; : 1-21, 2023 Apr 19.
Artículo en Inglés | MEDLINE | ID: mdl-37074167

RESUMEN

Pea proteins are widely used as a food ingredient, especially in sustainable food formulations. The seed itself consists of many proteins with different structures and properties that determine their structure-forming properties in food matrices, such as emulsions, foams, and gels. This review discusses the current insights into the structuring properties of pea protein mixtures (concentrates, isolates) and the resulting individual fractions (globulins, albumins). The structural molecular features of the proteins found in pea seeds are discussed and based on this information, different structural length scales relevant to foods are reviewed. The main finding of this article is that the different pea proteins are able to form and stabilize structural components found in foods such as air-water and oil-water interfaces, gels, and anisotropic structures. Current research reveals that each individual protein fraction has unique structure-forming properties and that tailored breeding and fractionation processes will be required to optimize these properties. Especially the use of albumins, globulins, and mixed albumin-globulins proved to be useful in specific food structures such as foams, emulsions, and self-coacervation, respectively. These new research findings will transform how pea proteins are processed and being used in novel sustainable food formulations in the future.

4.
Int J Mol Sci ; 24(16)2023 Aug 18.
Artículo en Inglés | MEDLINE | ID: mdl-37629113

RESUMEN

Although incurable pathologies associated with the formation of highly ordered fibrillar protein aggregates called amyloids have been known for about two centuries, functional roles of amyloids have been studied for only two decades. Recently, we identified functional amyloids in plants. These amyloids formed using garden pea Pisum sativum L. storage globulin and vicilin, accumulated during the seed maturation and resisted treatment with gastric enzymes and canning. Thus, vicilin amyloids ingested with food could interact with mammalian proteins. In this work, we analyzed the effects of vicilin amyloids on the fibril formation of proteins that form pathological amyloids. We found that vicilin amyloids inhibit the fibrillogenesis of these proteins. In particular, vicilin amyloids decrease the number and length of lysozyme amyloid fibrils; the length and width of ß-2-microglobulin fibrils; the number, length and the degree of clustering of ß-amyloid fibrils; and, finally, they change the structure and decrease the length of insulin fibrils. Such drastic influences of vicilin amyloids on the pathological amyloids' formation cause the alteration of their toxicity for mammalian cells, which decreases for all tested amyloids with the exception of insulin. Taken together, our study, for the first time, demonstrates the anti-amyloid effect of vicilin fibrils and suggests the mechanisms underlying this phenomenon.


Asunto(s)
Amiloide , Pisum sativum , Animales , Proteínas de Almacenamiento de Semillas , Insulina , Insulina Regular Humana , Mamíferos
5.
Int J Mol Sci ; 24(8)2023 Apr 21.
Artículo en Inglés | MEDLINE | ID: mdl-37108842

RESUMEN

Narrow-leafed lupin (NLL; Lupinus angustifolius L.) has multiple nutraceutical properties that may result from unique structural features of ß-conglutin proteins, such as the mobile arm at the N-terminal, a structural domain rich in α-helices. A similar domain has not been found in other vicilin proteins of legume species. We used affinity chromatography to purify recombinant complete and truncated (without the mobile arm domain, tß5 and tß7) forms of NLL ß5 and ß7 conglutin proteins. We then used biochemical and molecular biology techniques in ex vivo and in vitro systems to evaluate their anti-inflammatory activity and antioxidant capacity. The complete ß5 and ß7 conglutin proteins decreased pro-inflammatory mediator levels (e.g., nitric oxide), mRNA expression levels (iNOS, TNFα, IL-1ß), and the protein levels of pro-inflammatory cytokine TNF-α, interleukins (IL-1ß, IL-2, IL-6, IL-8, IL-12, IL-17, IL-27), and other mediators (INFγ, MOP, S-TNF-R1/-R2, and TWEAK), and exerted a regulatory oxidative balance effect in cells as demonstrated in glutathione, catalase, and superoxide dismutase assays. The truncated tß5 and tß7 conglutin proteins did not have these molecular effects. These results suggest that ß5 and ß7 conglutins have potential as functional food components due to their anti-inflammatory and oxidative cell state regulatory properties, and that the mobile arm of NLL ß-conglutin proteins is a key domain in the development of nutraceutical properties, making NLL ß5 and ß7 excellent innovative candidates as functional foods.


Asunto(s)
Lupinus , Lupinus/metabolismo , Suplementos Dietéticos
6.
Molecules ; 28(4)2023 Feb 07.
Artículo en Inglés | MEDLINE | ID: mdl-36838575

RESUMEN

Food allergy is a potentially life-threatening health concern caused by immunoglobulin E (IgE) antibodies that mistakenly recognize normally harmless food proteins as threats. Peanuts and tree nuts contain several seed storage proteins that commonly act as allergens. Glandless cottonseed, lacking the toxic compound gossypol, is a new food source. However, the seed storage proteins in cottonseed may act as allergens. To assess this risk, glandless cottonseed protein extracts were evaluated for IgE binding by peanut and tree nut allergic volunteers. ELISA demonstrated that 25% of 32 samples had significant binding to cottonseed extracts. Immunoblot analysis with pooled sera indicated that IgE recognized a pair of bands migrating at approximately 50 kDa. Excision of these bands and subsequent mass-spectrometric analysis demonstrated peptide matches to cotton C72 and GC72 vicilin and legumin A and B proteins. Further, in silico analysis indicated similarity of the cotton vicilin and legumin proteins to peanut vicilin (Ara h 1) and cashew nut legumin (Ana o 2) IgE-binding epitopes among others. The observations suggest both the cotton vicilin and legumin proteins were recognized by the nut allergic IgE, and they should be considered for future allergen risk assessments evaluating glandless cottonseed protein products.


Asunto(s)
Fabaceae , Hipersensibilidad a los Alimentos , Humanos , Nueces , Arachis/metabolismo , Aceite de Semillas de Algodón , Inmunoglobulina E , Alérgenos/química , Fabaceae/metabolismo , Proteínas de Almacenamiento de Semillas , Proteínas de Plantas/metabolismo , Antígenos de Plantas
7.
Curr Issues Mol Biol ; 44(7): 3100-3117, 2022 Jul 06.
Artículo en Inglés | MEDLINE | ID: mdl-35877438

RESUMEN

The aim of the study presented here was to determine if there is a correlation between the presence of specific protein domains within tree nut allergens or tree nut allergen epitopes and the frequency of bioactive fragments and the predicted susceptibility to enzymatic digestion in allergenic proteins from tree nuts of cashew (Anacardium occidentale), pecan (Carya illinoinensis), English walnut (Juglans regia) and pistachio (Pistacia vera) plants. These bioactive peptides are distributed along the length of the protein and are not enriched in IgE epitope sequences. Classification of proteins as bioactive peptide precursors based on the presence of specific protein domains may be a promising approach. Proteins possessing a vicilin, N-terminal family domain, or napin domain contain a relatively low occurrence of bioactive fragments. In contrast, proteins possessing the cupin 1 domain without the vicilin N-terminal family domain contain a relatively high total frequency of bioactive fragments and predicted release of bioactive fragments by the joint action of pepsin, trypsin, and chymotrypsin. This approach could be utilized in food science to simplify the selection of protein domains enriched for bioactive peptides.

8.
Molecules ; 27(10)2022 May 19.
Artículo en Inglés | MEDLINE | ID: mdl-35630727

RESUMEN

Vicilin has nutraceutical potential and different noteworthy medicative health-promoting biotic diversions, and it is remarkable against pathogenic microorganisms and insects. In this study, Vigna aconitifolia vicilin (VacV) has been identified and characterized from the seed of Vigna aconitifolia (Jacq.) Marechal (Moth beans). LC-MS/MS analysis of VacV provided seven random fragmented sequences comprising 238 residues, showing significant homology with already reported Vigna radiata vicilin (VraV). VacV was purified using ammonium sulfate precipitation (60%) followed by size exclusion chromatography on Hi-Load 16/60 Superdex 200 pg column and anion-exchange chromatography (Hi trap Q FF column). Purified VacV showed a major ~50 kDa band and multiple lower bands on 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) under both reduced and non-reduced conditions. After all, a three-dimensional molecular structure of VacV was predicted, which showed ß-sheeted molecular conformation similar to crystallographic structure of VraV. All Vicilins from V. aconitifolia and other plants were divided into six sub-groups by phylogenetic analysis, and VacV shared a high degree of similarity with vicilins of Vigna radiata, Pisum sativum, Lupinus albus, Cicer arietinum and Glycine max. Additionally, VacV (20 µg) has significant growth inhibition against different pathogenic bacteria along strong antifungal activity (50 µg). Likewise, VacV (3.0 mg) produced significant growth reduction in Rice Weevil Sitophilus oryzae larvae after 9 days compared with control. Furthermore, by using MMT assay, the cytotoxicity effect of VacV on the growth of HepG2 liver cancerous cells was tested. VacV showed cytotoxicity against the HepG-2 line and the acquired value was 180 µg after 48 h. Finally, we performed molecular docking against caspase-3 protein (PDB ID: 3DEI) for VacV bioactive receptor interface residues. Hence, our results reveal that VacV, has nutraceutical potential and moth beans can be used as a rich resource of functional foods.


Asunto(s)
Antiinfecciosos , Insecticidas , Vigna , Antibacterianos/análisis , Antiinfecciosos/análisis , Antiinfecciosos/farmacología , Cromatografía Liquida , Insecticidas/análisis , Insecticidas/farmacología , Simulación del Acoplamiento Molecular , Filogenia , Proteínas de Plantas/química , Proteínas de Almacenamiento de Semillas , Semillas/química , Espectrometría de Masas en Tándem
9.
Int J Mol Sci ; 22(6)2021 Mar 16.
Artículo en Inglés | MEDLINE | ID: mdl-33809562

RESUMEN

Adzuki seed ß-vignin, a vicilin-like globulin, has proven to exert various health-promoting biological activities, notably in cardiovascular health. A simple scalable enrichment procedure of this protein for further nutritional and functional studies is crucial. In this study, a simplified chromatography-independent protein fractionation procedure has been optimized and described. The electrophoretic analysis showed a high degree of homogeneity of ß-vignin isolate. Furthermore, the molecular features of the purified protein were investigated. The adzuki bean ß-vignin was found to have a native size of 146 kDa, and the molecular weight determined was consistent with a trimeric structure. These were identified in two main polypeptide chains (masses of 56-54 kDa) that are glycosylated polypeptides with metal binding capacity, and one minor polypeptide chain with a mass 37 kDa, wherein these features are absent. The in vitro analysis showed a high degree of digestibility of the protein (92%) and potential anti-inflammatory capacity. The results lay the basis not only for further investigation of the health-promoting properties of the adzuki bean ß-vignin protein, but also for a possible application as nutraceutical molecule.


Asunto(s)
Cromatografía/métodos , Proteínas de Plantas/genética , Vigna/química , Secuencia de Aminoácidos , Células CACO-2 , Fraccionamiento Químico , Harina , Globulinas/química , Humanos , Concentración de Iones de Hidrógeno , Inflamación/patología , Proteínas de Plantas/química , Proteínas de Plantas/aislamiento & purificación , Semillas/química , Solubilidad
10.
Clin Exp Allergy ; 50(5): 625-635, 2020 05.
Artículo en Inglés | MEDLINE | ID: mdl-32078204

RESUMEN

BACKGROUND: Food allergy to pea (Pisum sativum) has been rarely studied in children at the clinical and molecular levels. OBJECTIVE: To elucidate the allergenic relevance and diagnostic value of pea 7S globulin Pis s 1, nsLTP, and 2S albumins PA1 and PA2 in children. METHODS: Children with pea-specific IgE ≥ 0.35 kUA /L and clinical evidence of pea allergy or tolerance were included in the study. IgE binding against pea total protein extract, recombinant (r) rPis s 1, rPA1, rPA2, and natural nsLTP was analysed using IgE immunoblot/inhibition. Mediator release potency was investigated in passively sensitized rat basophil leukaemia (RBL) 2H3-cells. IgE binding to synthetic overlapping peptides of Pis s 1 was detected on multipeptide microarrays. RESULTS: 19 pea-sensitized children were included, 14 with doctors' diagnosed allergy and 5 with tolerance to pea (median age 3.5 and 4.5 years, respectively). 11/14 (78%) pea-allergic and 1/5 (20%) tolerant children were sensitized to Pis s 1. Under the reducing conditions of immunoblot analysis, IgE binding to rPA1 was negligible, sensitization to rPA2 and nsLTP undetectable. Compared to pea total protein extract, rPis s 1 displayed on average 58% IgE binding capacity and a 20-fold higher mediator release potency. Selected Pis s 1-related peptides displayed IgE binding in pea-allergic but not in pea-tolerant children. CONCLUSIONS AND CLINICAL RELEVANCE: In this study group, Pis s 1 is a major immunodominant allergen in pea-allergic children. Evidence for sensitization to nsLTP and 2S albumins was low but requires further verification with regard to conformational epitopes. Recombinant Pis s 1 and related peptides which were exclusively recognized by pea-allergic children may improve in vitro diagnosis of pea allergy once verified in prospective studies with larger study groups.


Asunto(s)
Alérgenos , Hipersensibilidad a los Alimentos , Inmunoglobulina E/inmunología , Pisum sativum , Adolescente , Alérgenos/química , Alérgenos/genética , Alérgenos/inmunología , Animales , Sitios de Unión , Niño , Preescolar , Femenino , Hipersensibilidad a los Alimentos/diagnóstico , Hipersensibilidad a los Alimentos/inmunología , Humanos , Lactante , Masculino , Pisum sativum/genética , Pisum sativum/inmunología , Proteínas de Plantas/química , Proteínas de Plantas/genética , Proteínas de Plantas/inmunología , Ratas
11.
J Sci Food Agric ; 100(13): 4657-4663, 2020 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-32270879

RESUMEN

Almond (Prunus dulcis) is not only widely used as a human food as a result of its flavor, nutrients, and health benefits, but it is also one of the most likely tree nuts to trigger allergies. Almond allergens, however, have not been studied as extensively as those of peanuts and other selected tree nuts. This review provides an update of the molecular properties of almond allergens to clarify some confusion about the identities of almond allergens and our perspective on characterizing putative almond allergens. At present, the following almond allergens have been designated by the World Health Organization/International Union of Immunological Societies Allergen Nomenclature Sub-Committee: Pru du 3 (a non-specific lipid transfer protein 1, nsLTP1), Pru du 4 (a profilin), Pru du 5 (60S acidic ribosomal protein 2), Pru du 6 (an 11S legumin known as prunin) and Pru du 8 (an antimicrobial protein with cC3C repeats). Besides, almond vicilin and almond γ-conglutin have been identified as food allergens, although further characterization of these allergens is still of interest. In addition, almond 2S albumin was reported as a food allergen as a result of the misidentification of Pru du 8. Two more almond proteins have been called allergens based on their sequence homology with known food allergens and their 'membership' in relevant protein families that contain allergens in many species. These include the pathogenesis related-10 protein (referred to as Pru du 1) and the thaumatin-like protein (referred to as Pru du 2). Almonds thus have five known food allergens and five more likely ones that need to be investigated further. Published 2020. This article is a U.S. Government work and is in the public domain in the USA.


Asunto(s)
Antígenos de Plantas/inmunología , Hipersensibilidad a los Alimentos/inmunología , Proteínas de Plantas/inmunología , Prunus dulcis/inmunología , Animales , Antígenos de Plantas/química , Antígenos de Plantas/genética , Humanos , Proteínas de Plantas/química , Proteínas de Plantas/genética , Prunus dulcis/química , Prunus dulcis/genética , Proteínas de Almacenamiento de Semillas/química , Proteínas de Almacenamiento de Semillas/genética , Proteínas de Almacenamiento de Semillas/inmunología
12.
J Sci Food Agric ; 100(7): 2889-2897, 2020 May.
Artículo en Inglés | MEDLINE | ID: mdl-32031252

RESUMEN

BACKGROUND: Alternaria tenuissima was isolated from infected fig fruit and molecularly identified by rRNA gene sequencing. The objective of the current work was to test the inhibitory effect of vicilin as a glycoprotein, isolated from chickpea, against the fungus A. tenuissima, isolated from fig fruit, in vitro and in situ, to estimate its potential action in controlling the growth of A. tenuissima in postharvest fig fruit. RESULTS: Chickpea vicilin is a glycoprotein composed of three subunits of 135, 210, and 230 kDa. The linear growth of A. tenuissima on the solid agar medium and in liquid media (at 25 °C) was markedly reduced by 44%, 66%, 77%, and 83% and 20%, 24%, 42%, and 62%, respectively in response to vicilin applications of 0.1, 0.2, 0.3, and 0.4 g L-1 . Chickpea vicilin (at 0.4 g L-1 ) totally prevented fungal conidia germination during 24 h of incubation at 25 °C. Electron microscope scanning of A. tenuissima subjected to chickpea vicilin showed hyphae swelling and conidia deformation. Treating post-harvest fig fruit, artificially infected with A. tenuissima, with chickpea vicilin (0.1-0.4 g L-1 ) restricted the disease severity to 15% against 55% in the positive control after 7 days storage. CONCLUSION: Vicilin can be considered a potent antifungal agent that can be used in preserving fig fruit for 7-14 days with minimum disease severity. © 2020 Society of Chemical Industry.


Asunto(s)
Alternaria/efectos de los fármacos , Ficus/microbiología , Fungicidas Industriales/farmacología , Proteínas de Almacenamiento de Semillas/farmacología , Alternaria/crecimiento & desarrollo , Alternaria/aislamiento & purificación , Cicer/química , Microbiología de Alimentos , Frutas/microbiología
14.
Biochem J ; 475(19): 3057-3071, 2018 10 09.
Artículo en Inglés | MEDLINE | ID: mdl-30181145

RESUMEN

Proteins belonging to cupin superfamily are known to have critical and diverse physiological functions. However, 7S globulins family, which is also a part of cupin superfamily, were undermined as only seed storage proteins. Structure determination of native protein - Vic_CAPAN from Capsicum annuum - was carried out, and its physiological functions were explored after purifying the protein by ammonium sulfate precipitation followed by size exclusion chromatography. The crystal structure of vicilin determined at 2.16 Šresolution revealed two monomers per asymmetric unit which are juxtaposed orthogonal with each other. Vic_CAPAN consists predominately of ß-sheets that folds to form a ß-barrel structure commonly called cupin fold. Each monomer of Vic_CAPAN consists of two cupin fold domains, N-terminal and C-terminal, which accommodate two different ligands. A bound ligand was identified at the C-terminal cupin fold in the site presumably conserved for metabolites in the crystal structure. The ligand was confirmed to be salicylic acid through mass spectrometric analysis. A copper-binding site was further observed near the conserved ligand-binding pocket, suggesting possible superoxide dismutase activity of Vic_CAPAN which was subsequently confirmed biochemically. Vicilins from other sources did not exhibit this activity indicating functional specificity of Vic_CAPAN. Discovery of bound salicylic acid, which is a known regulator of antioxidant pathway, and revelation of superoxide dismutase activity suggest that Vic_CAPAN has an important role during oxidative stress. As salicylic acid changes the redox state of cell, it may act as a downstream signal for various pathways involved in plant biotic and abiotic stress rescue.


Asunto(s)
Capsicum , Estrés Oxidativo/fisiología , Extractos Vegetales/química , Extractos Vegetales/metabolismo , Proteínas de Almacenamiento de Semillas/química , Proteínas de Almacenamiento de Semillas/metabolismo , Secuencia de Aminoácidos , Sitios de Unión/fisiología , Cristalización , Extractos Vegetales/genética , Estructura Secundaria de Proteína , Proteínas de Almacenamiento de Semillas/genética , Semillas
15.
Biosci Biotechnol Biochem ; 82(2): 285-291, 2018 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-29338640

RESUMEN

Cowpea seed ß-vignin, a vicilin-like globulin, proved to exert various health favourable effects, including blood cholesterol reduction in animal models. The need of a simple scalable enrichment procedure for further studies for tailored applications of this seed protein is crucial. A chromatography-independent fractionation method allowing to obtain a protein preparation with a high degree of homogeneity was used. Further purification was pursued to deep the molecular characterisation of ß-vignin. The results showed: (i) differing glycosylation patterns of the two constituent polypeptides, in agreement with amino acid sequence features; (ii) the seed accumulation of a gene product never identified before; (iii) metal binding capacity of native protein, a property observed only in few other legume seed vicilins.


Asunto(s)
Globulinas/química , Globulinas/metabolismo , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Vigna/química , Glicosilación , Metales/metabolismo , Semillas/química
17.
J Proteome Res ; 14(11): 4823-33, 2015 Nov 06.
Artículo en Inglés | MEDLINE | ID: mdl-26426307

RESUMEN

Coconut pollen, one of the major palm pollen grains is an important constituent among vectors of inhalant allergens in India and a major sensitizer for respiratory allergy in susceptible patients. To gain insight into its allergenic components, pollen proteins were analyzed by two-dimensional electrophoresis, immunoblotted with coconut pollen sensitive patient sera, followed by mass spectrometry of IgE reactive proteins. Coconut being largely unsequenced, a proteomic workflow has been devised that combines the conventional database-dependent analysis of tandem mass spectral data and manual de novo sequencing followed by a homology-based search for identifying the allergenic proteins. N-terminal acetylation helped to distinguish "b" ions from others, facilitating reliable sequencing. This led to the identification of 12 allergenic proteins. Cluster analysis with individual patient sera recognized vicilin-like protein as a major allergen, which was purified to assess its in vitro allergenicity and then partially sequenced. Other IgE-sensitive spots showed significant homology with well-known allergenic proteins such as 11S globulin, enolase, and isoflavone reductase along with a few which are reported as novel allergens. The allergens identified can be used as potential candidates to develop hypoallergenic vaccines, to design specific immunotherapy trials, and to enrich the repertoire of existing IgE reactive proteins.


Asunto(s)
Alérgenos/inmunología , Cocos/química , Proteínas de Plantas/aislamiento & purificación , Polen/inmunología , Hipersensibilidad Respiratoria/inmunología , Proteínas de Almacenamiento de Semillas/aislamiento & purificación , Acetilación , Alérgenos/química , Secuencia de Aminoácidos , Análisis por Conglomerados , Cocos/fisiología , Minería de Datos/estadística & datos numéricos , Electroforesis en Gel Bidimensional , Globulinas/química , Globulinas/inmunología , Globulinas/aislamiento & purificación , Humanos , Sueros Inmunes/química , Inmunoglobulina E/química , Anotación de Secuencia Molecular , Datos de Secuencia Molecular , Oxidorreductasas actuantes sobre Donantes de Grupo CH-CH/química , Oxidorreductasas actuantes sobre Donantes de Grupo CH-CH/inmunología , Oxidorreductasas actuantes sobre Donantes de Grupo CH-CH/aislamiento & purificación , Fosfopiruvato Hidratasa/química , Fosfopiruvato Hidratasa/inmunología , Fosfopiruvato Hidratasa/aislamiento & purificación , Proteínas de Plantas/química , Proteínas de Plantas/inmunología , Polen/química , Hipersensibilidad Respiratoria/sangre , Hipersensibilidad Respiratoria/fisiopatología , Proteínas de Almacenamiento de Semillas/química , Proteínas de Almacenamiento de Semillas/inmunología , Análisis de Secuencia de Proteína , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción
18.
Biopolymers ; 102(4): 335-43, 2014 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-24817604

RESUMEN

The objective of this study was to isolate antimicrobial peptides from Capsicum baccatum seeds and evaluate their antimicrobial activity and inhibitory effects against α-amylase. Initially, proteins from the flour of C. baccatum seeds were extracted in sodium phosphate buffer, pH 5.4, and precipitated with ammonium sulfate at 90% saturation. The D1 and D2 fractions were subjected to antifungal tests against the yeasts Saccharomyces cerevisiae, Candida albicans, Candida tropicalis, and Kluyveromyces marxiannus, and tested against α-amylases from Callosobruchus maculates and human saliva. The D2 fraction presented higher antimicrobial activity and was subjected to further purification and seven new different fractions (H1-H7) were obtained. Peptides in the H4 fraction were sequenced and the N-terminal sequences revealed homology with previously reported storage vicilins from seeds. The H4 fraction exhibited strong antifungal activity and also promoted morphological changes in yeast, including pseudohyphae formation. All fractions, including H4, inhibited mammalian α-amylase activity but only the H4 fraction was able to inhibit C. maculatus α-amylase activity. These results suggest that the fractions isolated from the seeds of C. baccatum can act directly in plant defenses against pathogens and insects.


Asunto(s)
Antifúngicos/farmacología , Capsicum/química , Péptidos/farmacología , Proteínas de Almacenamiento de Semillas/farmacología , Semillas/química , Levaduras/efectos de los fármacos , alfa-Amilasas/antagonistas & inhibidores , Secuencia de Aminoácidos , Animales , Antifúngicos/aislamiento & purificación , Cromatografía por Intercambio Iónico , Inhibidores Enzimáticos/farmacología , Humanos , Insectos , Pruebas de Sensibilidad Microbiana , Datos de Secuencia Molecular , Micología , Péptidos/química , Péptidos/aislamiento & purificación , Proteínas de Almacenamiento de Semillas/química , Proteínas de Almacenamiento de Semillas/aislamiento & purificación , Alineación de Secuencia , Levaduras/crecimiento & desarrollo , alfa-Amilasas/metabolismo
19.
Int J Biol Macromol ; 268(Pt 2): 131646, 2024 May.
Artículo en Inglés | MEDLINE | ID: mdl-38636765

RESUMEN

Plant-based food proteins are a promising choice for the preparation of nanoparticles (NPs) due to their high digestibility, low cost, and ability to interact with various compounds and nutrients. Moreover, nanoencapsulation offers a potential solution for protecting nutrients during processing and enhancing their bioavailability. This study aimed to develop and evaluate nanoparticles (NPs) based on legumin/vicilin (LV) proteins extracted from fava beans, with the goal of encapsulating and delivering a model nutraceutical compound, folic acid (FA). Specifically, NPs were self-assembled from LV proteins extracted from commercially available frozen fava beans using a pH-coacervation method with poloxamer 188 (P188) and chemically cross-linked with glutaraldehyde. Microscopy and spectroscopy studies were carried out on the empty and FA-loaded NPs in order to evaluate the particle morphology, size, size distribution, composition, mechanism of formation, impact of FA loading and release behavior. In vitro studies with Caco-2 cells also confirmed that the empty and FA-loaded nanoparticles were non-toxic. Thus, the LV-NPs are good candidates as food additives for the delivery and stabilization of nutrients as well as in drug delivery for the controlled release of therapeutics.


Asunto(s)
Preparaciones de Acción Retardada , Ácido Fólico , Nanopartículas , Poloxámero , Ácido Fólico/química , Humanos , Nanopartículas/química , Poloxámero/química , Células CACO-2 , Preparaciones de Acción Retardada/química , Liberación de Fármacos , Tamaño de la Partícula , Proteínas de Plantas/química , Portadores de Fármacos/química , Composición de Medicamentos
20.
J Agric Food Chem ; 71(6): 2990-2998, 2023 Feb 15.
Artículo en Inglés | MEDLINE | ID: mdl-36728846

RESUMEN

Peanut and tree-nut allergies are frequently comorbid for reasons not completely understood. Vicilin-buried peptides (VBPs) are an emerging family of food allergens whose conserved structural fold could mediate peanut/tree-nut co-allergy. Peptide microarrays were used to identify immunoglobulin E (IgE) epitopes from the N-terminus of the vicilin allergens Ara h 1, Ana o 1, Jug r 2, and Pis v 3 using serum from three patient diagnosis groups: monoallergic to either peanuts or cashew/pistachio, or dual allergic. IgE binding peptides were highly prevalent in the VBP domains AH1.1, AO1.1, JR2.1, and PV3.1, but not in AO1.2, JR2.2, JR2.3, and PV3.2 nor the unstructured regions. The IgE profiles did not correlate with diagnosis group. The structure of the VBPs from cashew and pistachio was solved using solution-NMR. Comparisons of structural features suggest that the VBP scaffold from peanuts and tree-nuts can support cross-reactivity. This may help understand comorbidity and cross-reactivity despite a distant evolutionary origin.


Asunto(s)
Anacardium , Arachis , Inmunoglobulina E , Juglans , Pistacia , Humanos , Alérgenos/química , Alérgenos/inmunología , Anacardium/química , Arachis/química , Inmunoglobulina E/inmunología , Juglans/química , Hipersensibilidad a la Nuez/diagnóstico , Nueces/química , Péptidos/química , Péptidos/inmunología , Pistacia/química , Reacciones Cruzadas
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