Division of Plant Science and Technology, University of Missouri, Columbia, MO, United States.
The Protein Facility of the Iowa State University Office of Biotechnology, Ames, IA, United States.
Methods Enzymol. 2022;676:325-345. doi: 10.1016/bs.mie.2022.07.001. Epub 2022 Jul 29.
The salt-soluble globulins, glycinins (11S globulin), and β-conglycinins (7S globulin), are the most abundant seed proteins of soybean seeds. Together, these two groups of proteins account for 60-70% of total soybean seed proteins. Proteomic assessment of the less abundant soybean seed proteins using general isolation protocols is challenging due to the overwhelming abundance of storage proteins. Development of a simple, fast, and inexpensive method to remove most storage proteins from a seed extract will significantly enhance the study of the nonabundant proteins within seeds. We have developed two simple methods for the depletion of abundant seed proteins resulting in the enrichment of low abundance proteins from soybean seeds. Here, we provide a detailed procedure for the isolation, separation, identification, and quantification of low abundance seed proteins of soybean.
盐溶性球蛋白,即甘氨酸(11S 球蛋白)和 β-伴球蛋白(7S 球蛋白),是大豆种子中含量最丰富的种子蛋白。这两类蛋白共同占大豆种子总蛋白的 60-70%。由于大量存在的贮藏蛋白,使用常规分离方法对含量较少的大豆种子蛋白进行蛋白质组学评估具有挑战性。开发一种简单、快速且廉价的方法从种子提取物中去除大部分贮藏蛋白,将显著促进对种子内低丰度蛋白的研究。我们开发了两种从大豆种子中去除丰富的种子蛋白的简单方法,从而富集低丰度蛋白。在此,我们提供了一种详细的大豆低丰度种子蛋白分离、分离、鉴定和定量的程序。