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Proteomics Analysis of Embryo and Endosperm from Mature Common Buckwheat Seeds

机译:成熟普通荞麦种子胚和胚乳的蛋白质组学分析

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We used proteomics analysis to generate the profiles of proteins in the endosperm and embryo of common buckwheat grains. These differentially expressed proteins are potentially involved in seed metabolism. Extractions were done by trichloroacetic acid(TCA) precipitation. The resulting proteins were separated using SDS-PAGE coupled to LC-ESI-Q/TOF-MS/MS. This allowed us to detect and identify 67 proteins with isoforms, making this the most inclusive protein profile. The proteins were determined to befunctionally involved in the central metabolic pathway of the seed, with metabolic interest being reflected in the occurrence of a tissue-specific enzyme balance. For a case in point, we found a tissue-specific and subcellular compartment-specific isoform of granule-bound starch synthase 1 in the chloroplast/amyloplast. This provided proteomic verification of the presence of a distinct regulatory mechanism for the biosynthesis of glycan and starch, which produce amylase and amylopectin. Furthermore, several previously characterized allergenic proteins such as 11S and 13S globulin seed storage protein were acknowledged in our seed samples, thus representing the potential for proteomics techniques that survey food sources for any incidence of allergens.This protein profile of common buckwheat grain is a new avenue for understanding its seed physiology in dormant stage as well as suggesting commercial applications for the buckwheat industry as buckwheat flour.
机译:我们使用蛋白质组学分析来生成常见荞麦谷物的胚乳和胚胎中蛋白质的概况。这些差异表达的蛋白质可能参与种子代谢。通过三氯乙酸(TCA)沉淀进行提取。使用偶联至LC-ESI-Q / TOF-MS / MS的SDS-PAGE分离所得蛋白质。这使我们能够检测和鉴定67个具有同工型的蛋白质,这使它成为最具包容性的蛋白质谱。确定蛋白质在功能上参与种子的中央代谢途径,代谢兴趣反映在组织特异性酶平衡的发生上。例如,我们在叶绿体/淀粉质体中发现了颗粒结合淀粉合酶1的组织特异性和亚细胞室特异性亚型。这提供了蛋白质组学的验证,证明了产生淀粉酶和支链淀粉的聚糖和淀粉生物合成的独特调节机制的存在。此外,我们的种子样品中还确认了几种先前表征的变应原蛋白,例如11S和13S球蛋白种子贮藏蛋白,从而代表了蛋白质组学技术对食物来源进行调查以查明任何变应原的可能性。了解其休眠期种子生理的途径,并建议将荞麦粉作为荞麦工业的商业应用。

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