解读小麦粉的免疫原性潜能:美国小麦Butte 86的盐溶性蛋白质组参考图。

IF 2.1 3区 生物学 Q3 BIOCHEMICAL RESEARCH METHODS Proteome Science Pub Date : 2020-08-01 eCollection Date: 2020-01-01 DOI:10.1186/s12953-020-00164-6
Susan B Altenbach, Han-Chang Chang, Annamaria Simon-Buss
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引用次数: 5

摘要

背景:在复杂的小麦粉蛋白质组中,面筋蛋白因其在决定小麦粉面团的功能特性及其在乳糜泻和食物过敏等人类健康状况中的重要作用而引起了人们的广泛关注。然而,某些非麸质蛋白也会引发免疫反应,但可能在面粉中含量较低,或被总蛋白制备的二维凝胶中更丰富的麸质蛋白所掩盖。方法:用稀盐溶液优先提取面粉中的非面筋蛋白,用双向凝胶电泳分离。用胰蛋白酶或凝乳胰蛋白酶裂解后,用串联质谱法鉴定了173个凝胶点的蛋白。利用RNA干扰抑制特定谷蛋白群的转基因小麦品系,估计了盐溶性蛋白部分中谷蛋白的携带量。结果:共鉴定出57种不同类型的非面筋蛋白,其中14种为已知或疑似免疫原性蛋白。18个凝胶点的优势蛋白为面筋蛋白。其中一些还含有非麸质蛋白。通过RNA干扰去除omega-1,2麦胶蛋白的转基因品系的盐溶性蛋白分析表明,某些omega-1,2麦胶蛋白在盐溶性部分中大量存在,并掩盖了相对少量的β -淀粉酶和蛋白质二硫异构酶。相比之下,对缺乏α -麦胶蛋白的转基因品系的分析表明,甘油醛-3磷酸脱氢酶是一种中等含量的蛋白质,可与几种α -麦胶蛋白共迁移。结论:在这项研究中,我们构建了美国Butte 86小麦面粉的非面筋蛋白部分的蛋白质组学图谱,补充了之前为同一品种开发的总面粉蛋白质组学图谱。了解面粉中低丰度蛋白的特性,以及被一些主要面筋蛋白隐藏在二维凝胶中的蛋白的特性,对于评估小麦粉的免疫原性潜力,以及确定在未来的基因编辑实验中应该针对哪些小麦蛋白来降低小麦粉的免疫原性潜力的研究至关重要。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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Deciphering the immunogenic potential of wheat flour: a reference map of the salt-soluble proteome from the U.S. wheat Butte 86.

Background: Within the complex wheat flour proteome, the gluten proteins have attracted most of the attention because of their importance in determining the functional properties of wheat flour doughs and their roles in human health conditions such as celiac disease and food allergies. However, certain non-gluten proteins also trigger immunological responses but may be present in flour in low amounts or obscured by the more abundant gluten proteins in two-dimensional gels of total protein preparations.

Methods: Non-gluten proteins were preferentially extracted from the flour with a dilute salt solution and separated by two-dimensional gel electrophoresis. Proteins in 173 gel spots were identified by tandem mass spectrometry after cleavage with trypsin or chymotrypsin. Transgenic wheat lines in which specific groups of gluten proteins were suppressed by RNA interference were used to estimate the amount of carry-over of gluten proteins in the salt-soluble protein fraction.

Results: Fifty-seven different types of non-gluten proteins were identified, including 14 types that are known or suspected immunogenic proteins. The predominant proteins in 18 gel spots were gluten proteins. Some of these also contained non-gluten proteins. Analysis of the salt-soluble proteins from a transgenic line in which omega-1,2 gliadins were eliminated by RNA interference indicated that certain omega-1,2 gliadins were present in large amounts in the salt-soluble fraction and obscured relatively small amounts of beta-amylase and protein disulfide isomerase. In comparison, analysis of a transgenic line in which alpha gliadins were absent revealed that glyceraldehyde-3 phosphate dehydrogenase was a moderately abundant protein that co-migrated with several alpha gliadins.

Conclusions: In this study, we constructed a proteomic map of the non-gluten protein fraction of wheat flour from the US wheat Butte 86 that complements a proteomic map of the total flour proteins developed previously for the same cultivar. Knowing the identities of low abundance proteins in the flour as well as proteins that are hidden by some of the major gluten proteins on two-dimensional gels is critical for studies aimed at assessing the immunogenic potential of wheat flour and determining which wheat proteins that should be targeted in future gene editing experiments to reduce the immunogenic potential of wheat flour.

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来源期刊
Proteome Science
Proteome Science 生物-生化研究方法
CiteScore
2.90
自引率
0.00%
发文量
17
审稿时长
4.5 months
期刊介绍: Proteome Science is an open access journal publishing research in the area of systems studies. Proteome Science considers manuscripts based on all aspects of functional and structural proteomics, genomics, metabolomics, systems analysis and metabiome analysis. It encourages the submissions of studies that use large-scale or systems analysis of biomolecules in a cellular, organismal and/or environmental context. Studies that describe novel biological or clinical insights as well as methods-focused studies that describe novel methods for the large-scale study of any and all biomolecules in cells and tissues, such as mass spectrometry, protein and nucleic acid microarrays, genomics, next-generation sequencing and computational algorithms and methods are all within the scope of Proteome Science, as are electron topography, structural methods, proteogenomics, chemical proteomics, stem cell proteomics, organelle proteomics, plant and microbial proteomics. In spite of its name, Proteome Science considers all aspects of large-scale and systems studies because ultimately any mechanism that results in genomic and metabolomic changes will affect or be affected by the proteome. To reflect this intrinsic relationship of biological systems, Proteome Science will consider all such articles.
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