A label-free shotgun proteomics analysis of macadamia nut

Rost, Johanna; Muralidharan, Sridevi; Lee, Nanju Alice

doi:10.1016/j.foodres.2019.108838

Cited by 19 publications

(20 citation statements)

References 56 publications

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“…A crude extract of macadamia proteins was obtained following a well-established protocol, yielding an acceptable efficiency above 90%, which facilitated its lyophilization and prolonged storage. The extracted proteins revealed after SDS-PAGE and Coomassie staining (Figure ) a range of molecular masses from 10 to 70 kDa, which corresponded well to previous results . A significant difference was noticed when polypeptides were separated by SDS-PAGE under reducing or nonreducing conditions.…”

Section: Resultssupporting

confidence: 57%

Unbiased Immunization Strategy Yielding Specific Nanobodies against Macadamia Allergen of Vicilin-like Protein for Immunoassay Development

Wang

et al. 2021

J. Agric. Food Chem.

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Macadamia nut contains important food allergens that potentially cause allergic reactions with severe adverse effects in infants and adults. Reliable and accurate detection of macadamia is critical to avoid allergic reactions. However, knowledge on macadamia allergen is scarce and a reliable detection method has not been reported, yet. In this study, an unbiased immunization and selection strategy was employed to select nanobodies (Nbs) recognizing specifically macadamia allergen, as well as to establish a detection method to unveil a macadamia protein contamination. An alpaca was immunized with a crude protein extract of macadamia followed by construction of a Nb library from its lymphocytes. The panning and screening of this immune Nb repertoire resulted in the selection of six target-specific Nbs. Nb-mediated immuno-capturing combined with mass spectrometry allowed us to identify the target as the macadamia vicilin-like antimicrobial peptides 2–3 (MiAMP2), a novel food allergenic protein abbreviated as Mac i 1. Later on, an immunoassay of a heterologous sandwich ELISA method based on the selected Nb-pairs was established, providing a linear response in the range of 0.442–2,800 μg/mL and with a limit of detection of 27.1 ng/mL. The dedicated immunoassay has been verified by detecting the antigen spiked in food samples. Our study provided evidence for the successful application of the unprejudiced strategy to retrieve Nbs against a priori undefined macadamia allergen. These target-specific Nbs were used to design a highly reliable and effective immunoassay.

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Section: Resultssupporting

confidence: 57%

Unbiased Immunization Strategy Yielding Specific Nanobodies against Macadamia Allergen of Vicilin-like Protein for Immunoassay Development

Wang

et al. 2021

J. Agric. Food Chem.

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“…In addition to antimicrobial properties, these seed storage proteins are homologous to vicilin 7S globulins and have F I G U R E 2 Chromosomal distribution of duplicated genes of M. jansenii, generated using SynVisio. The parallel horizontal lines represent the 14 pseudo-molecules of M. jansenii genome, with connected ribbons representing the duplicated genes been identified as putative allergens (Rost et al, 2016(Rost et al, , 2020. A cDNA sequence, from M. integrifolia, encoding these proteins, MiAMP-2, has been reported to contain four repeat segments, with each segment comprised of cysteine rich motifs (C-X-X-X-C-[10 to12] X-C-X-X-X-C), where X is any other amino acid residue.…”

Section: Discussionmentioning

confidence: 99%

The genome of the endangered Macadamia jansenii displays little diversity but represents an important genetic resource for plant breeding

et al. 2021

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Macadamia, a recently domesticated expanding nut crop in the tropical and subtropical regions of the world, is one of the most economically important genera in the diverse and widely adapted Proteaceae family. All four species of Macadamia are rare in the wild with the most recently discovered, M. jansenii, being endangered. The M. jansenii genome has been used as a model for testing sequencing methods using a wide range of long read sequencing techniques. Here, we report a chromosome level genome assembly, generated using a combination of Pacific Biosciences sequencing and Hi-C, comprising 14 pseudo-molecules, with a N50 of 52 Mb and a total genome assembly size of 758 Mb of which 56% is repetitive. Completeness assessment revealed that the assembly covered À97.1% of the conserved single copy genes. Annotation predicted 31,591 protein coding genes and allowed the characterization of genes encoding biosynthesis of cyanogenic glycosides, fatty acid metabolism, and anti-microbial proteins. Re-sequencing of seven other genotypes confirmed low diversity and low heterozygosity within this endangered species. Important morphological characteristics of this species such as small tree size and high kernel recovery suggest that M. jansenii is an important source of these commercial traits for breeding. As a member of a small group of families that are sister to the core eudicots, this high-quality genome also provides a key resource for evolutionary and comparative genomics studies.

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“…However, recent advancement of LC-MS/MS-based methods mainly known as shotgun proteomics or discovery proteomics can be essential for this purpose. This platform provides highly accurate identification and quantitation of many known and unknown plant allergens and allergen-like proteins with their posttranslational modified forms ( Rahman et al, 2021 , Rost et al, 2020 , Spiric et al, 2017 ). Only a handful of gel-based and/or shotgun proteomic analyses have been performed to catalogue the total proteome and allergen proteins from lentil seed samples.…”

Section: Methodologies For Identification and Quantification Of Lenti...mentioning

confidence: 99%

Lentil allergens identification and quantification: An update from omics perspective

Halima

Najar

Wahab

et al. 2022

Food Chemistry: Molecular Sciences

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A label-free shotgun proteomics analysis of macadamia nut

Cited by 19 publications

References 56 publications

Unbiased Immunization Strategy Yielding Specific Nanobodies against Macadamia Allergen of Vicilin-like Protein for Immunoassay Development

Unbiased Immunization Strategy Yielding Specific Nanobodies against Macadamia Allergen of Vicilin-like Protein for Immunoassay Development

The genome of the endangered Macadamia jansenii displays little diversity but represents an important genetic resource for plant breeding

Lentil allergens identification and quantification: An update from omics perspective

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