Proteins are responsible for most intracellular functions, which they perform as part of higher-order molecular complexes, located within defined subcellular niches. Localization is both dynamic and context specific and mislocalization underlies a multitude of diseases. It is thus vital to be able to measure the components of higher-order protein complexes and their subcellular location dynamically in order to fully understand cell biological processes. Here, we review the current range of highly complementary approaches that determine the subcellular organization of the proteome. We discuss the scale and resolution at which these approaches are best employed and the caveats that should be taken into consideration when applying them. We also look to the future and emerging technologies that are paving the way for a more comprehensive understanding of the functional roles of protein isoforms, which is essential for unraveling the complexities of cell biology and the development of disease treatments.

蛋白质负责大多数细胞内功能，它们作为高级分子复合物的一部分发挥作用，位于确定的亚细胞生态位内。定位既是动态的又是特定于环境的，定位错误是多种疾病的基础。因此，为了充分了解细胞生物过程，能够动态测量高阶蛋白质复合物的成分及其亚细胞位置至关重要。在这里，我们回顾了当前确定蛋白质组亚细胞组织的高度互补的一系列方法。我们讨论了这些方法最适合采用的规模和分辨率，以及应用它们时应考虑的注意事项。我们还展望未来和新兴技术，这些技术为更全面地了解蛋白质亚型的功能作用铺平了道路，这对于揭示细胞生物学的复杂性和疾病治疗的发展至关重要。