An improved MALDI-TOF mass spectrometry procedure and a novel DNA marker for identifying over-expressed Bx7 glutenin protein subunit in wheat

Wang, Ke; Islam, Shahidul; Ma, Jing; Anwar, Masood; Chen, Jing; Yan, Yueming; Appels, R.; Ma, Wujun

doi:10.1111/hrd2.00069

Cited by 3 publications

(2 citation statements)

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“…A more recent analysis of a gluten food ingredient, coupled with fractionation to enrich gluten proteins also allowed identification of several hundred gluten-specific peptides (Martínez-Esteso et al, 2016). Due to the complexity of gluten, a large proportion of the current MS analysis carried out on gluten has been focused to profile HMW glutenin subunits initially (Lagrain et al, 2013; Wang et al, 2014). …”

Section: Introductionmentioning

confidence: 99%

Comprehensive Proteomic Profiling of Wheat Gluten Using a Combination of Data-Independent and Data-Dependent Acquisition

et al. 2017

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Wheat is the most important food crop in the world, the unique physiochemical properties of wheat gluten enabling a diverse range of food products to be manufactured. However, genetic and environmental factors affect the technological properties of gluten in unpredictable ways. Although newer proteomic methods have the potential to offer much greater levels of information, it is the older gel-based methods that remain most commonly used to identify compositional differences responsible for the variation in gluten functionality, in part due to the nature of their primary sequences. A combination of platforms were investigated for comprehensive gluten profiling: a QTOF with a data independent schema, which incorporated ion mobility (DIA-IM-MS) and a data dependent acquisition (DDA) workflow using a linear ion trap quadrupole (LTQ) instrument. In conjunction with a manually curated gluten sequence database a total of 2736 gluten peptides were identified with only 157 peptides identified by both platforms. These data showed 127 and 63 gluten protein accessions to be inferred with a minimum of one and three unique peptides respectively. Of the 63 rigorously identified proteins, 26 were gliadin species (4 ω-, 14 α-, and 8 γ-gliadins) and 37 glutenins (including 29 LMW glutenin and 8 HMW glutenins). Of the HMW glutenins, three were 1Dx type and five were 1Bx type illustrating the challenge of unambiguous identification of highly polymorphic proteins without cultivar specific gene sequences. The capacity of the platforms to sequence longer peptides was crucial to achieving the number of identifications, the combination of QTOF-LTQ technology being more important than extraction method to obtain a comprehensive profile. Widespread glutamine deamidation, a post-translational modification, was observed adding complexity to an already highly polymorphic mixture of proteins, with numerous insertions, deletions and substitutions. The data shown is the most comprehensive and detailed proteomic profile of gluten to date.

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Section: Introductionmentioning

confidence: 99%

Comprehensive Proteomic Profiling of Wheat Gluten Using a Combination of Data-Independent and Data-Dependent Acquisition

et al. 2017

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“…Eagles et al [23] found the greatest HMW-GS polymorphism in the Glu-B1 locus, in which over 30 alleles could be distinguished. The relationship between the presence of specific alleles in HMW-GS is extensively described in the literature [24][25][26][27][28]. In general, the data found in the literature suggest that an allele of one subunit has a higher negative impact on the dough energy than alleles of two subunits [21,25,27].…”

Section: Discussionmentioning

confidence: 99%

Composition of High Molecular Weight Glutenin Subunits in Polish Common Wheat Cultivars (Triticum aestivum L.)

Filip

2018

Journal of Food Quality

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The main goal of our study was to present research data on genes encoding high molecular weight glutenin subunits (HMW-GS) associated with high flour bread-making quality. This is the leading research objective in our institute in the area of wheat gluten in cultivars that have not been studied so far in that respect, but which can potentially be a valuable source of new information. Identification and characterization of high molecular weight glutenin subunits (HMW-GS) were performed using sequencing and SDS-PAGE and STS-PCR methods. Genes located in the vicinity of the Glu-1 locus have been identified and characterized in 28 Polish cultivars of Triticum aestivum. The results were then analyzed using the following computer programs: Finch TV, BLAST, MEGA 4, Molecular Imager® Gel Doc™ XR, and Quantity One software (Bio-Rad). Three alleles (a, b, c) have been identified in the Glu-A1 locus, 6 alleles (a, b, c, d, e, k) in the Glu-B1 locus, and 2 alleles (a, d) in Glu-D1 using the SDS-PAGE method. The amplification of specific HMW-GS sequences generated one product of 450 bp in 1Dx5 in 13 cultivars of old wheat and of 435 bp in 1Dx2 in 15 cultivars. The amplification products of primers for 1Dy10 and 1Dy12 genes were 422 bp and 552 bp in size, respectively.

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Introgression of an expressed HMW 1Ay glutenin subunit allele into bread wheat cv. Lincoln increases grain protein content and breadmaking quality without yield penalty

Roy

Islam

et al. 2019

Theor Appl Genet

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An improved MALDI-TOF mass spectrometry procedure and a novel DNA marker for identifying over-expressed Bx7 glutenin protein subunit in wheat

Cited by 3 publications

References 21 publications

Comprehensive Proteomic Profiling of Wheat Gluten Using a Combination of Data-Independent and Data-Dependent Acquisition

Comprehensive Proteomic Profiling of Wheat Gluten Using a Combination of Data-Independent and Data-Dependent Acquisition

Composition of High Molecular Weight Glutenin Subunits in Polish Common Wheat Cultivars (Triticum aestivum L.)

Introgression of an expressed HMW 1Ay glutenin subunit allele into bread wheat cv. Lincoln increases grain protein content and breadmaking quality without yield penalty

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