Project description:This SuperSeries is composed of the following subset Series: GSE15686: Meta-transcriptome analysis of a natural wheat sourdough ecosystem during a 10-day spontaneous laboratory fermentation (I) GSE15691: Meta-transcriptome analysis of a natural spelt sourdough ecosystem during a 10-day spontaneous laboratory fermentation (I) GSE15692: Meta-transcriptome analysis of a natural spelt sourdough ecosystem during a 10-day spontaneous laboratory fermentation (II) GSE15693: Meta-transcriptome analysis of a natural wheat sourdough ecosystem during a 10-day spontaneous laboratory fermentation (II) Refer to individual Series
Project description:LC-MS/MS-based LFQ proteomics of 150 wheat full-kernel flours from five different wheat species (common wheat, spelt, durum wheat, emmer, einkorn) grown at three different locations.
Project description:Amylase/trypsin-inhibitors (ATIs) are putative triggers of nonceliac gluten sensitivity, but contents of ATIs in different wheat species were not available. Therefore, the predominant ATIs 0.19 + 0.53, 0.28, CM2, CM3, and CM16 in eight cultivars each of common wheat, durum wheat, spelt, emmer, and einkorn grown under the same environmental conditions were quantitated by targeted liquid chromatography-tandem mass spectrometry (LC−MS/MS) and stable isotope dilution assays using specific marker peptides as internal standards. The results were compared to a label-free untargeted LC−MS/MS analysis, in which protein concentrations were determined by intensity based absolute quantitation. Both approaches yielded similar results. Spelt and emmer had higher ATI contents than common wheat, with durum wheat in between. Only three of eight einkorn cultivars contained ATIs in very low concentrations. The distribution of ATI types was characteristic for hexaploid, tetraploid, and diploid wheat species and suitable as species-specific fingerprint. The results point to a better tolerability of einkorn for NCGS patients, because of very low total ATI contents.
Project description:An untargeted liquid chromatography tandem mass spectrometry method was used to analyze the content of peptides with celiac disease (CD) active epitope in the five wheat species common wheat, spelt, durum wheat, emmer and einkorn. In total, 494 peptides with CD active epitope were identified. Relevant differences between the species concerning their CD immunoractive potential based on the distribution of CD-active epitopes and relative quantities of peptides with CD-active epitope were demonstrated.
Project description:In this study, we present a novel hybrid data-independent acquisition LC-MS approach, combining QconCAT technology with short microflow LC gradients and DIA and apply the method towards the quantitative proteome analysis of ATI extracts from wheat flour. The novel method is fast, robust and reproducible and provides accurate QconCAT-based absolute quantification of major ATI proteins while simultaneously quantifying the non-targeted proteome by DIA-based label-free quantification. Using a set of 120 samples of full kernel flour derived from 60 varieties of common wheat grown in replication, we assess the repeatability and accuracy of the workflow for the quantification of ATI at the peptide and protein level. Applying the method to analyze different wheat species (i.e. common wheat, spelt, durum wheat, emmer and einkorn) and comparing the results to published data, we validate inter-laboratory and cross-methodology reproducibility of ATI quantification, which are essential in the context of large-scale breeding projects. Additionally, we apply our workflow to assess environmental effects on ATI expression, analyzing ATI content and proteome of wheat species grown at different locations. Finally, we explore the potential of combining QconCAT-based absolute quantification with DIA-based untargeted proteome analysis and LFQ for the generation of new hypotheses or assay development.
Project description:High-molecular-weight glutenin subunits (HMW-GS) play an important role for the baking quality of wheat. The ancient wheats emmer and spelt differ in their HMW-GS pattern compared to modern common wheat and this might be one reason for their comparatively poor baking quality. The aim of this study was to elucidate similarities and differences in the amino acid sequences of two 1Bx HMW-GS of common wheat, spelt and emmer. First, the sodium dodecyl polyacrylamide gel electrophoresis (SDS-PAGE) system was optimized to separate common wheat, spelt and emmer Bx6 and Bx7 from other HMW-GS (e.g., 1Ax and 1By) in high concentrations. The in-gel digests of the Bx6 and Bx7 bands were analyzed by untargeted LC-MS/MS experiments revealing different UniProtKB accessions in spelt and emmer compared to common wheat. The HMW-GS Bx6 and Bx7, respectively, of emmer and spelt showed differences in the amino acid sequences compared to those of common wheat. The identities of the peptide variations were confirmed by targeted LC-MS/MS. These peptides can be used to differentiate between Bx6 and Bx7 of spelt and emmer and Bx6 and Bx7 of common wheat. The findings should help to increase the reliability and curation status of wheat protein databases and to understand the effects of protein structure on the functional properties.
2020-01-15 | PXD016342 | panorama
Project description:Assembly of Triticum aestivum cv. 'Spelt' as part of the 10+ Wheat Genomes Project.
Project description:Rye, wheat and barley contain gluten, proteins that trigger immune-mediated inflammation of the small intestine in people with coeliac disease (CD). The only treatment for CD is a lifelong gluten-free diet. To be classified as gluten-free by the World Health Organisation the gluten content must be below 20 mg/kg, but Australia has a more rigorous standard of no detectable gluten and not made from wheat, barley, rye or oats. The purpose of this study was to devise an LC-MS/MS method to detect rye in food. An MS-based assay could overcome some of the limitations of current immunoassays, wherein antibodies often show cross-reactivity and lack specificity due to the diversity of gluten proteins in commercial food and the homology between rye and wheat gluten isoforms. Comprehensive proteomic analysis of 20 rye cultivars originating from 12 countries enabled the identification of a panel of candidate rye-specific peptide markers. The peptide markers were assessed in 16 cereal and pseudo-cereal grains, and in 10 breakfast cereals and 7 snacks foods. Spelt flour was contaminated with rye at a level of 2% and trace levels of rye were found in a breakfast cereal that based on its labelled ingredients should be gluten-free.
2019-07-25 | PXD013459 | Pride
Project description:Analysis of natural wheat and spelt sourdough ecosystem during a 10-day spontaneous laboratory fermentation