Mapping Coeliac Toxic Motifs in the Prolamin Seed Storage Proteins of Barley, Rye, and Oats Using a Curated Sequence Database

A - Papers appearing in refereed journals

Daly, M., Bromilow, S. N., Nitride, C., Shewry, P. R., Gethings, L. A. and Mills, E. N. C 2020. Mapping Coeliac Toxic Motifs in the Prolamin Seed Storage Proteins of Barley, Rye, and Oats Using a Curated Sequence Database. Frontiers in Nutrition. 7, p. 87. https://doi.org/10.3389/fnut.2020.00087

AuthorsDaly, M., Bromilow, S. N., Nitride, C., Shewry, P. R., Gethings, L. A. and Mills, E. N. C
Abstract

Wheat gluten, and related prolamin proteins in rye, barley and oats cause the immune-mediated gluten intolerance syndrome, coeliac disease. Foods labelled as gluten-free which can be safely consumed by coeliac patients, must not contain gluten above a level of 20 mg/Kg. Current immunoassay methods for detection of gluten can give conflicting results and may underestimate levels of gluten in foods. Mass spectrometry methods have great potential as an orthogonal method, but require curated protein sequence databases to support method development. The GluPro database has been updated to include avenin-like sequences from bread wheat (n = 685; GluPro v1.1) and genes from the sequenced wheat genome (n = 699; GluPro v 1.2) and Triticum turgidum ssp durum (n = 210; GluPro v 2.1). Companion databases have been developed for prolamin sequences from barley (n = 64; GluPro v 3.0), rye (n = 41; GluPro v 4.0), and oats (n = 27; GluPro v 5.0) and combined to provide a complete cereal prolamin database, GluPro v 6.1 comprising 1,041 sequences. Analysis of the coeliac toxic motifs in the curated sequences showed that they were absent from the minor avenin-like proteins in bread and durum wheat and barley, unlike the related avenin proteins from oats. A comparison of prolamin proteins from the different cereal species also showed α- and γ-gliadins in bread and durum wheat, and the sulphur poor prolamins in all cereals had the highest density of coeliac toxic motifs. Analysis of ion-mobility mass spectrometry data for bread wheat (cvs Chinese Spring and Hereward) showed an increased number of identifications when using the GluPro v1.0, 1.1 and 1.2 databases compared to the limited number of verified sequences bread wheat sequences in reviewed UniProt. This family of databases will provide a basis for proteomic profiling of gluten proteins from all the gluten containing cereals and support identification of specific peptide markers for use in development of new methods for gluten quantitation based on coeliac toxic motifs found in all relevant cereal species.

KeywordsGluten; Sequence Database; Barley; Rye; Oats; Coeliac disease; Wheat
Year of Publication2020
JournalFrontiers in Nutrition
Journal citation7, p. 87
Digital Object Identifier (DOI)https://doi.org/10.3389/fnut.2020.00087
Open accessPublished as ‘gold’ (paid) open access
FunderBiotechnology and Biological Sciences Research Council
Funder project or codeDFW - Designing Future Wheat - Work package 2 (WP2) - Added value and resilience
Publisher's version
Output statusPublished
Publication dates
Print17 Jul 2020
Publication process dates
Accepted12 May 2020
PublisherFrontiers Media SA
ISSN2296-861X

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