MSU-DOE Plant Research Lab and Plant Biology Department, Michigan State University, East Lansing, MI, USA.
Methods Mol Biol. 2022;2378:279-289. doi: 10.1007/978-1-0716-1732-8_18.
Excessive accumulation of unfolded or misfolded proteins in the endoplasmic reticulum (ER) leads to a potentially cytotoxic condition known as the ER stress. Upon ER stress, cells initiate a homeostatic response called unfolded protein response (UPR) to assist proper folding the unfolded or misfolded proteins. Proteomics have been broadly used in plants with Liquid Chromatography coupled to tandem MS (LC-MS/MS) technologies. LC-MS/MS techniques have also been a great tool for studies of posttranslational modifications (PTMs). Here we describe our protocol of a fast method for large amount of seedling treatment and collection for UPR study in Arabidopsis thaliana and the preparation of total proteins for proteomic analysis.
内质网（ER）中未折叠或错误折叠蛋白质的过度积累会导致一种潜在的细胞毒性状态，称为内质网应激。内质网应激后，细胞会启动一种称为未折叠蛋白反应（UPR）的体内平衡反应，以协助未折叠或错误折叠蛋白质的正确折叠。蛋白质组学在植物中已广泛应用，与液相色谱串联质谱（LC-MS/MS）技术相结合。LC-MS/MS 技术也是研究翻译后修饰（PTMs）的重要工具。在这里，我们描述了一种快速方法的协议，用于大量幼苗处理和收集，以进行拟南芥 UPR 研究，并准备用于蛋白质组学分析的总蛋白质。
