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Advances in the Biology of Seed and Vegetative Storage Proteins Based on Two-Dimensional Electrophoresis Coupled to Mass Spectrometry

机译:二维电泳与质谱联用的种子和营养贮藏蛋白生物学研究进展

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摘要

Seed storage proteins play a fundamental role in plant reproduction and human nutrition. They accumulate during seed development as reserve material for germination and seedling growth and are a major source of dietary protein for human consumption. Storage proteins encompass multiple isoforms encoded by multi-gene families that undergo abundant glycosylations and phosphorylations. Two-dimensional electrophoresis (2-DE) is a proteomic tool especially suitable for the characterization of storage proteins because of their peculiar characteristics. In particular, storage proteins are soluble multimeric proteins highly represented in the seed proteome that contain polypeptides of molecular mass between 10 and 130 kDa. In addition, high-resolution profiles can be achieved by applying targeted 2-DE protocols. 2-DE coupled with mass spectrometry (MS) has traditionally been the methodology of choice in numerous studies on the biology of storage proteins in a wide diversity of plants. 2-DE-based reference maps have decisively contributed to the current state of our knowledge about storage proteins in multiple key aspects, including identification of isoforms and quantification of their relative abundance, identification of phosphorylated isoforms and assessment of their phosphorylation status, and dynamic changes of isoforms during seed development and germination both qualitatively and quantitatively. These advances have translated into relevant information about meaningful traits in seed breeding such as protein quality, longevity, gluten and allergen content, stress response and antifungal, antibacterial, and insect susceptibility. This review addresses progress on the biology of storage proteins and application areas in seed breeding using 2-DE-based maps.
机译:种子贮藏蛋白在植物繁殖和人类营养中起着基本作用。它们在种子发育过程中积累,作为发芽和幼苗生长的储备材料,是人类食用蛋白质的主要来源。贮藏蛋白包括由多基因家族编码的多种同工型,它们经历大量的糖基化和磷酸化。二维电泳(2-DE)是一种蛋白质组学工具,由于其独特的特性而特别适合表征存储蛋白。特别地,贮藏蛋白是在种子蛋白质组中高度代表的可溶性多聚体蛋白,其包含分子量在10至130kDa之间的多肽。此外,可以通过应用有针对性的2-DE协议来实现高分辨率配置文件。传统上,将2-DE与质谱(MS)结合使用是众多植物中多种贮藏蛋白生物学研究的首选方法。基于2-DE的参考图谱在许多关键方面为我们对存储蛋白的知识的发展做出了决定性的贡献,包括鉴定同工型和定量其相对丰度,鉴定磷酸化同工型以及评估其磷酸化状态以及动态变化。定性和定量分析种子发育和萌发过程中的同工型。这些进展已转化为有关种子育种中有意义特征的相关信息,例如蛋白质质量,寿命,面筋和过敏原含量,应激反应以及抗真菌,抗菌和昆虫敏感性。这篇综述探讨了基于2-DE的图谱在种子育种中贮藏蛋白生物学及其应用领域的进展。

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