Takahashi Daisuke, Nakayama Takato, Miki Yushi, Kawamura Yukio, Uemura Matsuo
United Graduate School of Agricultural Sciences, Iwate University, 3-18-8 Ueda, Morioka, Iwate, 020-8550, Japan.
Methods Mol Biol. 2014;1166:159-70. doi: 10.1007/978-1-4939-0844-8_13.
Plasma membrane is the primary determinant of freezing tolerance in plants because of its central role in freeze-thaw cycle. Changes in the plasma membrane proteins 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核心作用。质膜蛋白的变化一直是植物低温驯化的主要研究领域之一。为了获得质膜蛋白质组的全面概况及其在低温驯化过程中的变化,本文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以及一种基于质谱的鸟枪法蛋白质组学方法,该方法使用纳米液相色谱 - 串联质谱分析质膜蛋白。所获得的蛋白质组学结果进一步应用于无标记蛋白质半定量分析。