Extraction of wheat endosperm proteins for proteome analysis
Extraction of wheat endosperm proteins for proteome analysis
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DOI：
10.1016/j.jchromb.2006.11.047
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发表时间：
2007-04-15
影响因子：
3
通讯作者：
Tanaka, Charlene K.
中科院分区：
文献类型：
--
作者：
Hurkman, William J.;Tanaka, Charlene K.
小麦胚乳总蛋白提取物被广泛用于分析含量丰富的醇溶蛋白和谷蛋白。在这篇综述中，最流行的总胚乳提取方法进行了比较，其有效性的蛋白质组覆盖。总胚乳提取物的缺点是蛋白质丰度的巨大动态范围限制了低丰度蛋白质的检测、定量和鉴定。蛋白质分级分离对于提高蛋白质组覆盖率是非常宝贵的，因为它降低了样品的复杂性，同时富集了特定类别的丰度较低的蛋白质。广泛的技术可用于分离蛋白质亚群。连续提取是一种特别适合于小麦胚乳蛋白亚分级的方法，因为它利用了不同种类胚乳蛋白的特定溶解性。该方法有效地将高丰度的麦醇溶蛋白和麦谷蛋白与低得多的清蛋白和球蛋白分离。组织匀浆的亚细胞分级分离是分离膜和细胞器用于功能分析的经典技术。这种方法适用于定义与淀粉体相关的生化过程，淀粉体是胚乳中在淀粉合成和储存中起作用的专门细胞器。亚蛋白质组分离，结合2-DE和蛋白质鉴定，提供了一个强大的方法来确定胚乳蛋白质的组成，并提供新的见解细胞功能。(c)2006 Elsevier B. V.保留所有权利。
Total protein extracts of wheat endosperm are widely used for the analysis of the highly abundant gliadins and glutenins. In this review, the most popular total endosperm extraction methods are compared for their effectiveness in proteome coverage. A drawback of total endosperm extracts is that the enormous dynamic range of protein abundance limits the detection, quantification, and identification of low abundance proteins. Protein fractionation is invaluable for improving proteome coverage, because it reduces sample complexity while enriching for specific classes of less abundant proteins. A wide array of techniques is available for isolating protein subpopulations. Sequential extraction is a method particularly suited for subfractionation of wheat endosperm proteins, because it takes advantage of the specific solubility properties of the different classes of endosperm proteins. This method effectively separates the highly abundant gliadins and glutenins from the much less abundant albumins and globulins. Subcellular fractionation of tissue homogenates is a classical technique for isolating membranes and organelles for functional analysis. This approach is suitable for defining the biochemical processes associated with amyloplasts, specialized organelles in the endosperm that function in the synthesis and storage of starch. Subproteome fractionation, when combined with 2-DE and protein identification, provides a powerful approach for defining endosperm protein composition and providing new insights into cellular functions. (c) 2006 Elsevier B.V. All rights reserved.