Dynamic Cell Imaging, Faculty of Biology, Bielefeld University, Bielefeld, Germany.
Methods Mol Biol. 2024;2832:115-132. doi: 10.1007/978-1-0716-3973-3_8.
Proteins often show alterations in their subcellular localization with changing environmental conditions; transcription factors enter the nucleus or are actively removed from the nucleus; some even bind to endo-membranes by conditional membrane anchors; and other proteins and mRNA arrange in RNA granules. These are some examples of the complex regulation of subcellular localization, which often depends on posttranslational modifications and is triggered by environmental stressors. The challenge is the precise identification of the compartments, the quantitative analysis of proteins, which reside in multiple compartments, and their transport dynamics. Therefore, appropriate compartment markers and routines for a reproducible quantitative workflow are required.
蛋白质在环境条件改变时通常会表现出亚细胞定位的改变;转录因子进入核内或被主动从核内去除;有些甚至通过条件性膜锚定结合到内体膜上;其他蛋白质和 mRNA 排列在 RNA 颗粒中。这些是亚细胞定位复杂调控的一些例子,这种调控通常依赖于翻译后修饰,并由环境胁迫触发。挑战在于准确识别隔室,对驻留在多个隔室中的蛋白质进行定量分析,以及分析它们的运输动力学。因此,需要适当的隔室标记物和可重复的定量工作流程例程。