Rapid separation of seed gliadins by reversed-phase ultra performance liquid chromatography (RP-UPLC) and its application in wheat cultivar and germplasm identification

Han, Caixia; Lu, Xaiobing; Yu, Zitong; Li, Xiaohui; Ma, Wujun; Yan, Yueming

doi:10.1080/09168451.2014.998618

Cited by 16 publications

(12 citation statements)

References 41 publications

(41 reference statements)

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“…Our recent studies have demonstrated that using RP‐UPLC as a new method for wheat storage protein separation and characterization has clear advantages in terms of separation time, resolution, reproducibility and reagent consumption compared with reverse phase high‐performance liquid chromatography (RP‐HPLC) . In this study, the dynamic accumulation patterns of gliadins, HMW‐GS and LMW‐GS from 15 DPA to grain maturity under well‐watered and water deficit conditions were detected by RP‐UPLC.…”

Section: Resultsmentioning

confidence: 87%

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Effects of water deficit on breadmaking quality and storage protein compositions in bread wheat (Triticum aestivum L.)

et al. 2018

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Section: Resultsmentioning

confidence: 87%

“…Gliadins and glutenins were extracted from three biological replicates and separated by RP‐UPLC according to recent reports of Yu et al ., Han et al . and Yan et al .…”

Section: Methodsmentioning

confidence: 99%

Effects of water deficit on breadmaking quality and storage protein compositions in bread wheat (Triticum aestivum L.)

et al. 2018

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“…HPLC, UPLC, and MALDI-TOF technologies have been applied to separate wheat prolamins [67][68][69]. Procedures for sequentially extracting and recovering protein fractions from small flour samples were previously reported [35].…”

Section: Discussionmentioning

confidence: 99%

Characterising avenin-like proteins (ALPs) from albumin/globulin fraction of wheat grains by RP-HPLC, SDS-PAGE, and MS/MS peptides sequencing

Zhang

Juhász

et al. 2020

BMC Plant Biol

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Background: Wheat grain avenin-like proteins (ALPs) belong to a recently discovered class of wheat grain storage protein. ALPs in wheat grains not only have beneficial effects on dough quality but also display antifungal activities, which is a novel observation for wheat storage proteins. Previous studies have shown that ALPs are likely present in the albumin/globulin fractions of total protein extract from wheat flour. However, the accumulation characteristics of these ALPs in the mature wheat grain remains unknown. Results: In the present study, a total of 13 ALPs homologs were isolated and characterized in the albumin/globulin fractions of the wheat protein extract. A combination of multiple techniques including RP-HPLC, SDS-PAGE, MALDI-TOF and peptide sequencing were used for accurate separation and identification of individual ALP homolog. The C-terminal TaALP-by-4AL/7DS, TaALP-by-4AL/7AS/7DS, TaALP-bx/4AL/7AS/7DS, TaALP-ay-7DS, TaALP-ay-4AL, TaALPax-4AL, TaALP-ax-7AS, and TaALP-ax-7DS, were separated as individual protein bands from wheat flour for the first time. These unique ALPs peptides were mapped to the latest wheat genome assembly in the IWGSC database. The characteristic defence related proteins present in albumin and globulin fractions, such as protein disulfide-isomerase (PDI), grain softness protein (GSP), alpha-amylase inhibitors (AAIs) and endogenous alpha-amylase/subtilisin inhibitor were also found to co-segregate with these identified ALPs, avenin-3 and α-gliadins. The molecular weight range and the electrophoresis segregation properties of ALPs were characterised in comparison with the proteins containing the tryp_alpha_amyl domain (PF00234) and the gliadin domain (PF13016), which play a role in plant immunity and grain quality. We examined the phylogenetic relationships of the AAIs, GSP, avenin-3, α-gliadins and ALPs, based on the alignment of their functional domains. MALDI-TOF profiling indicated the occurrence of certain post-translations modifications (PTMs) in some ALP subunits. Conclusions: We reported for the first time the complete profiling of ALPs present in the albumin/globulin fractions of wheat grain protein extracts. We concluded that majority of the ALPs homologs are expressed in wheat grains. We found clear evidence of PTMs in several ALPs peptides. The identification of both gliadin domain (PF13016) and Tryp_alpha_amyl domain (PF00234) in the mature forms of ALPs highlighted the multiple functional properties of ALPs in grain quality and disease resistance.

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“…Current high-throughput proteomic methodologies based on LC-MS can be grouped into shotgun and targeted proteomics, or based on their data acquisition mode e.g., Data Dependent Acquisition (DDA) and the more recent Data Independent Acquisition (DIA ) ( Figure 5 ). The advances in LC separation and MS instrumentation are providing more and more high analytical reproducibility, speed in acquisition, high-resolution and sensitivity up to femtomoles [ 57 , 58 , 59 ]. The improvement of these aspects, coupled with sample preparation protocols [ 60 ] and advanced bioinformatic tools [ 61 ], may be translated into higher plant proteome coverage, including post-translational modifications (PTMs) and PPIs [ 62 ]; PTMs identification often requires specific enrichment steps, like in the case of phosphorylation or glycosylation, which complexify sample preparation procedure as well as protein quantitation [ 63 ].…”

Section: Omic Technologies In the Plant World: From Genomics To Mementioning

confidence: 99%

Large Scale Proteomic Data and Network-Based Systems Biology Approaches to Explore the Plant World

et al. 2018

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The investigation of plant organisms by means of data-derived systems biology approaches based on network modeling is mainly characterized by genomic data, while the potential of proteomics is largely unexplored. This delay is mainly caused by the paucity of plant genomic/proteomic sequences and annotations which are fundamental to perform mass-spectrometry (MS) data interpretation. However, Next Generation Sequencing (NGS) techniques are contributing to filling this gap and an increasing number of studies are focusing on plant proteome profiling and protein-protein interactions (PPIs) identification. Interesting results were obtained by evaluating the topology of PPI networks in the context of organ-associated biological processes as well as plant-pathogen relationships. These examples foreshadow well the benefits that these approaches may provide to plant research. Thus, in addition to providing an overview of the main-omic technologies recently used on plant organisms, we will focus on studies that rely on concepts of module, hub and shortest path, and how they can contribute to the plant discovery processes. In this scenario, we will also consider gene co-expression networks, and some examples of integration with metabolomic data and genome-wide association studies (GWAS) to select candidate genes will be mentioned.

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Rapid separation of seed gliadins by reversed-phase ultra performance liquid chromatography (RP-UPLC) and its application in wheat cultivar and germplasm identification

Cited by 16 publications

References 41 publications

Effects of water deficit on breadmaking quality and storage protein compositions in bread wheat (Triticum aestivum L.)

Effects of water deficit on breadmaking quality and storage protein compositions in bread wheat (Triticum aestivum L.)

Characterising avenin-like proteins (ALPs) from albumin/globulin fraction of wheat grains by RP-HPLC, SDS-PAGE, and MS/MS peptides sequencing

Large Scale Proteomic Data and Network-Based Systems Biology Approaches to Explore the Plant World

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