Laboratory of Cellular and Structural Biology, The Rockefeller University , New York, NY, USA.
Laboratory of Mass Spectrometry and Gaseous Ion Chemistry, The Rockefeller University , New York, NY, USA.
J Cell Biol. 2023 Jun 5;222(6). doi: 10.1083/jcb.202209062. Epub 2023 Mar 15.
Subcellular fractionation in combination with mass spectrometry-based proteomics is a powerful tool to study localization of key proteins in health and disease. Here we offered a reliable and rapid method for mammalian cell fractionation, tuned for such proteomic analyses. This method proves readily applicable to different cell lines in which all the cellular contents are accounted for, while maintaining nuclear and nuclear envelope integrity. We demonstrated the method's utility by quantifying the effects of a nuclear export inhibitor on nucleoplasmic and cytoplasmic proteomes.
亚细胞组分分离与基于质谱的蛋白质组学相结合是研究健康和疾病中关键蛋白质定位的有力工具。在这里,我们提供了一种可靠且快速的哺乳动物细胞组分分离方法,针对此类蛋白质组学分析进行了优化。该方法适用于不同的细胞系,可确保所有细胞内容物都被分离出来,同时保持核和核膜的完整性。我们通过定量分析核输出抑制剂对核质和细胞质蛋白质组的影响,证明了该方法的实用性。