Mass spectrometry-based proteomics for abiotic stress studies
Mass spectrometry-based proteomics for abiotic stress studies
复制标题
基于质谱的蛋白质组学用于非生物胁迫研究
DOI：
10.1016/j.tplants.2021.12.013
复制
发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Yingfang Zhu
中科院分区：
文献类型：
--
作者：
Leelyn Chong;Yingfang Zhu
激酶测定连接磷酸蛋白质组学（KALIP）和通用磷酸蛋白质组学（UP）工作流程是用于研究非生物胁迫信号网络的基于质谱（MS）的磷酸蛋白质组学技术。KALIP包括在体外进行同位素标记的磷酸化反应和在体内筛选磷酸化肽/底物池。对于UP，盐酸胍（GdnHCl）用于破坏细胞壁，然后应用甲醇-氯仿沉淀以去除干扰。这两种策略的结论与随后的分析富集磷酸肽互补MS技术，以确定磷酸肽丰度的变化和蛋白质磷酸化位点，发现新的信号转导通路中的组件。
Kinase assay linked phosphoproteomics (KALIP) and universal phosphoproteomics (UP) workflows are mass spectrometry (MS)-based phosphoproteomics technologies utilized for studying abiotic stress signaling networks. KALIP consists of executing isotope-labeled phosphorylation reactions in vitro and screening in vivo phosphorylated peptides/pools of substrates. For UP, guanidine hydrochloride (GdnHCl) is used to break down the cell wall before methanol-chloroform precipitation is applied to remove interferences. Both strategies conclude with subsequent analyses of enriched phosphopeptides by complementary MS techniques to determine phosphopeptide abundance changes and protein phosphorylation sites to uncover novel signaling components in the pathway.