Evaluation of protein extraction methods for in-depth proteome analysis of narrow-leafed lupin (Lupinus angustifolius) seeds

Tahmasian, Arineh; Broadbent, James A.; Juhász, Angéla; Nye-Wood, Mitchell G.; Le, Thao T.; Bose, Utpal; Colgrave, Michelle L.

doi:10.1016/j.foodchem.2021.130722

Cited by 13 publications

(6 citation statements)

References 38 publications

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“…The relative abundances of the γ1-conglutin peptides were significantly higher in D2-D5, and D7, whilst the γ2 specific peptides were found at significantly higher levels in the D4, D6, and D7 cultivars, this elevation may implicate the compensation for the suppressed β-conglutin levels across these domesticated lines, for ensuring the overall protein quantity within the seed, a phenomenon which has been previously reported for other seeds such as ultra-low-gluten barley cultivars (46) and transgenic soybean seeds (47). These NLL cultivars with lower allergenic β-conglutin content and higher bioactive γ-conglutin levels may offer enhanced health benefits; however, the lupin seed proteome also includes several thousand moderate or low abundant proteins (19). For deeper understanding of the proteome composition and rebalancing mechanisms a comprehensive investigation of the proteomewide alterations is essential.…”

Section: Gamma-conglutin Protein Familymentioning

confidence: 75%

“…A recently optimized urea-based protocol was used for the efficient extraction of the proteins from lupin seeds (19). Briefly, 800 µL of urea buffer (8 M urea, 2% (w/v) DTT, 1% (v/v) protease inhibitor cocktail in 0.1 M Tris-HCl; pH 8.2) was added to 20 mg of defatted lupin varieties (n = 3).…”

Section: Protein Extraction and Protein Estimationmentioning

confidence: 99%

“…The conglutin proteins identified at 1% FDR from the combined database searching were imported into Skyline 2 https://www.iedb.org/ software (v19.1.0.193) and subjected to in silico digestion. The ProteinPilot group files from the discovery proteomics data collected from lupin samples previously in our laboratory (19) and the combined search from the present study were used to build a BiblioSpec library containing peptides with a confidence value of > 0.95 (1% FDR). The library matched tryptic peptides (allowing one missed cleavage) between 8 to 30 amino acids in length with no variable modifications retained for the preliminary analyses (278 peptides).…”

Section: Targeted Assay Development and Peptide Specificity Analysismentioning

confidence: 99%

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Evaluation of the Major Seed Storage Proteins, the Conglutins, Across Genetically Diverse Narrow-Leafed Lupin Varieties

et al. 2022

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Lupin seeds have an excellent nutritional profile, including a high proportion of protein and dietary fiber. These qualities make lupin seeds an ideal candidate to help meet the growing global demand for complementary sources of protein. Of consequence to this application, there are nutritional and antinutritional properties assigned to the major lupin seed storage proteins—referred to as α-, β-, δ- and γ-conglutins The variation in the abundance of these protein families can impact the nutritional and bioactive properties of different lupin varieties. Hence, exploring the conglutin protein profiles across a diverse range of lupin varieties will yield knowledge that can facilitate the selection of superior genotypes for food applications or lupin crop improvement. To support this knowledge generation, discovery proteomics was applied for the identification of the 16 known conglutin subfamilies from 46 domestic and wild narrow-leafed lupin (NLL) genotypes. Consequently, the diversity of abundance of these proteins was evaluated using liquid chromatography–multiple reaction monitoring-mass spectrometry (LC–MRM-MS). This comparative study revealed a larger variability for the β- and δ-conglutin content across the lines under study. The absence/lower abundance of the β2- to β6-conglutin subfamilies in a subset of the domesticated cultivars led to substantially lower overall levels of the allergenic β-conglutin content in these NLLs, for which the elevation of the other conglutin families were observed. The diversity of the conglutin profiles revealed through this study—and the identification of potential hypoallergenic genotypes—will have great significance for lupin allergic consumers, food manufactures as well as grain breeders through the future development of lupin varieties with higher levels of desirable bioactive proteins and lower allergen content.

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Section: Gamma-conglutin Protein Familymentioning

confidence: 75%

Section: Protein Extraction and Protein Estimationmentioning

confidence: 99%

Section: Targeted Assay Development and Peptide Specificity Analysismentioning

confidence: 99%

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Evaluation of the Major Seed Storage Proteins, the Conglutins, Across Genetically Diverse Narrow-Leafed Lupin Varieties

et al. 2022

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“…It is interesting to highlight the wide biological properties that this group of bioactive peptides shows depending on the peptide studied, including antihypertensive, antioxidant, antiproliferative, anti-inflammatory, apiaceous, hypocholesteremia, antithrombotic and mineral chelator functions [ 40 , 41 , 42 , 43 , 44 , 53 ]. On the other hand, phenolic compounds, including flavonoids (anthocyanins, flavanols, catechins, gallo catechins, etc.…”

Section: Sources Of Nutraceuticals With Biological Valuementioning

confidence: 99%

Nutraceuticals and Food-Grade Lipid Nanoparticles: From Natural Sources to a Circular Bioeconomy Approach

Blanco-Llamero

Fonseca

Durazzo³

et al. 2022

Foods

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Nutraceuticals have gained increasing attention over the last years due to their potential value as therapeutic compounds formulated from natural sources. For instance, there is a wide range of literature about the cardioprotective properties of omega-3 lipids and the antioxidant value of some phenolic compounds, which are related to antitumoral activity. However, the value of nutraceuticals can be limited by their instability under gastric pH and intestinal fluids, their low solubility and absorption. That is why encapsulation is a crucial step in nutraceutical design. In fact, pharmaceutical nanotechnology improves nutraceutical stability and bioavailability through the design and production of efficient nanoparticles (NPs). Lipid nanoparticles protect the bioactive compounds from light and external damage, including the gastric and intestinal conditions, providing a retarded delivery in the target area and guaranteeing the expected therapeutic effect of the nutraceutical. This review will focus on the key aspects of the encapsulation of bioactive compounds into lipid nanoparticles, exploring the pharmaceutical production methods available for the synthesis of NPs containing nutraceuticals. Moreover, the most common nutraceuticals will be discussed, considering the bioactive compounds, their natural source and the described biological properties.

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“…In some cases, unrooted phenetic trees were built from the distance matrix calculated according to the Jaccard index on all spots from the analyzed cultivars using neighbor-joining algorithms. Recently, label-free shotgun proteomics has been used to investigate quali-quantitative differences between root, seed, or fruit tissues from different pea ( Meisrimler et al., 2017 ), grape ( Carpentieri et al., 2019 ), quinoa ( Galindo-Lujan et al., 2021 ), and narrow-leafed lupin ( Tahmasian et al., 2022 ) accessions, respectively. Statistical approaches generated peculiar heat-maps reporting quantitative protein profiles of cultivars, whose dendrogram from unsupervised hierarchical cluster analysis described the similarity of accessions, whereas PCA score biplot diagrams of protein percentage data among them illustrated phenotype relationships.…”

Section: Introductionmentioning

confidence: 99%

Investigating phenotypic relationships in persimmon accessions through integrated proteomic and metabolomic analysis of corresponding fruits

et al. 2023

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Together with phenological and genomic approaches, gel-based and label-free proteomic as well metabolomic procedures were separately applied to plants to highlight differences between ecotypes, to estimate genetic variability within/between organism populations, or to characterize specific mutants/genetically modified lines at metabolic level. To investigate the possible use of tandem mass tag (TMT)-based quantitative proteomics in the above-mentioned contexts and based on the absence of combined proteo-metabolomic studies on Diospyros kaki cultivars, we here applied integrated proteomic and metabolomic approaches to fruits from Italian persimmon ecotypes with the aim to characterize plant phenotypic diversity at molecular level. We identified 2255 proteins in fruits, assigning 102 differentially represented components between cultivars, including some related to pomological, nutritional and allergenic characteristics. Thirty-three polyphenols were also identified and quantified, which belong to hydroxybenzoic acid, flavanol, hydroxycinnamic acid, flavonol, flavanone and dihydrochalcone sub-classes. Heat-map representation of quantitative proteomic and metabolomic results highlighted compound representation differences in various accessions, whose elaboration through Euclidean distance functions and other linkage methods defined dendrograms establishing phenotypic relationships between cultivars. Principal component analysis of proteomic and metabolomic data provided clear information on phenotypic differences/similarities between persimmon accessions. Coherent cultivar association results were observed between proteomic and metabolomic data, emphasizing the utility of integrating combined omic approaches to identify and validate phenotypic relationships between ecotypes, and to estimate corresponding variability and distance. Accordingly, this study describes an original, combined approach to outline phenotypic signatures in persimmon cultivars, which may be used for a further characterization of other ecotypes of the same species and an improved description of nutritional characteristics of corresponding fruits.

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Evaluation of protein extraction methods for in-depth proteome analysis of narrow-leafed lupin (Lupinus angustifolius) seeds

Cited by 13 publications

References 38 publications

Evaluation of the Major Seed Storage Proteins, the Conglutins, Across Genetically Diverse Narrow-Leafed Lupin Varieties

Evaluation of the Major Seed Storage Proteins, the Conglutins, Across Genetically Diverse Narrow-Leafed Lupin Varieties

Nutraceuticals and Food-Grade Lipid Nanoparticles: From Natural Sources to a Circular Bioeconomy Approach

Investigating phenotypic relationships in persimmon accessions through integrated proteomic and metabolomic analysis of corresponding fruits

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