富膜蛋白质组分析的增溶和制备方法比较

Y. Tsujimoto, N. Taguchi, Kiyoo Hirooka, Naohiro Tomari, Akiko Okami, T. Nishikawa, Y. Nakazawa, Kunihiko Watanabe, H. Matsui, Yoshihiro Yamamoto
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引用次数: 2

摘要

由于膜蛋白的溶解问题,利用二维电泳(2-DE)对膜蛋白进行蛋白质组学分析仍然不足。此外,已知核酸和脂质会干扰2-DE的轮廓,通常需要去除这些物质。在本研究中,使用Bio-X公司的三种试剂去除蛋白质中的这些干扰物质。在上样前,用试剂在30℃下处理样品30-60 min。用这些试剂进行预处理可以实现高分辨率的蛋白质分离。在这里,我们描述了样品预处理如何使用试剂改善从大肠杆菌和酿酒酵母中提取的蛋白质的蛋白质组学图。去脂增强了膜蛋白与细胞的溶解性和分离性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Comparison of solubilization and preparation procedures for membrane-enriched proteome analysis
Proteome analysis of membrane proteins by two-dimentional electrophoresis (2-DE) is still insufficient due to the problem of membrane proteins solubilization. Additionally, nucleic acids and lipids are known to interfere with the profile on 2-DE, and removal of these substances is often required. In this study, three reagents from Bio-X Inc. were used to remove these interfering substances from protein. Prior to loading, samples were treated with the reagents for 30-60 min at 30°C. Pretreatment with the reagents led to high-resolution separations of proteins. Here, we describe how sample pretreatment using the reagents improved proteomic maps of proteins extracted from Escherichia coli and Saccharomyces cerevisiae. To remove lipids enhanced the solubility and separation of membrane proteins from cells.
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