United Graduate School of Agricultural Sciences, Faculty of Agriculture, Iwate University, Morioka, Iwate, Japan.
Graduate School of Science & Engineering, Saitama University, Saitama, Japan.
Methods Mol Biol. 2020;2156:171-186. doi: 10.1007/978-1-0716-0660-5_13.
Plasma membrane is the primary determinant of freezing tolerance in plants because of its central role in freeze-thaw cycle. Changes in plasma membrane protein composition have been one of the major research areas in plant cold acclimation. To obtain comprehensive profiles of the plasma membrane proteomes and their changes during the cold acclimation process, a plasma membrane purification method using a dextran-polyethylene glycol two polymer system and a mass spectrometry-based shotgun proteomics method using nano-LC-MS/MS for the plasma membrane proteins are described. The proteomic results obtained are further applied to label-free protein semiquantification.
质膜是植物抗冻性的主要决定因素，因为它在冻融循环中起着核心作用。质膜蛋白组成的变化是植物冷驯化的主要研究领域之一。为了获得质膜蛋白质组及其在冷驯化过程中变化的综合图谱，描述了一种使用葡聚糖-聚乙二醇双聚合物系统的质膜纯化方法和一种基于纳升液相色谱-串联质谱(nano-LC-MS/MS)的用于质膜蛋白质的鸟枪法蛋白质组学方法。进一步将获得的蛋白质组学结果应用于无标记蛋白质半定量。
