Subcellular proteomics
Subcellular proteomics
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DOI：
10.1038/s43586-021-00029-y
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发表时间：
2021-04-29
期刊：
影响因子：
--
通讯作者：
Lilley, Kathryn S.
中科院分区：
文献类型：
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作者：
Christopher, Josie A.;Stadler, Charlotte;Lilley, Kathryn S.
真核细胞被划分为亚细胞壁龛，包括膜结合细胞器和无膜细胞器。蛋白质定位于这些壁龛来实现它们的功能，使离散的生物过程同步发生。蛋白质在生态位之间的动态移动对于细胞过程至关重要，如信号传导、生长、增殖、运动和程序性细胞死亡，而引起异常蛋白质定位的突变与多种疾病有关。确定蛋白质在不同细胞状态和细胞类型中的位置，以及蛋白质在扰动后如何重新定位，对于理解它们的功能、相关的细胞过程和与它们的错误定位相关的病理非常重要。在这篇引物中，我们介绍了用于确定亚细胞结构中蛋白质的位置、分布和丰度的主要空间蛋白质组学方法。这些技术包括荧光成像、蛋白质接近标记、细胞器纯化和全细胞生化分离。我们描述了它们的工作流程、数据输出和在探索不同细胞生物学场景中的应用，并讨论了它们的主要局限性。最后，我们描述了新兴技术，并确定了需要技术创新的领域，以便更好地表征空间蛋白质组。
The eukaryotic cell is compartmentalized into subcellular niches, including membrane-bound and membrane-less organelles. Proteins localize to these niches to fulfil their function, enabling discreet biological processes to occur in synchrony. Dynamic movement of proteins between niches is essential for cellular processes such as signalling, growth, proliferation, motility and programmed cell death, and mutations causing aberrant protein localization are associated with a wide range of diseases. Determining the location of proteins in different cell states and cell types and how proteins relocalize following perturbation is important for understanding their functions, related cellular processes and pathologies associated with their mislocalization. In this Primer, we cover the major spatial proteomics methods for determining the location, distribution and abundance of proteins within subcellular structures. These technologies include fluorescent imaging, protein proximity labelling, organelle purification and cell-wide biochemical fractionation. We describe their workflows, data outputs and applications in exploring different cell biological scenarios, and discuss their main limitations. Finally, we describe emerging technologies and identify areas that require technological innovation to allow better characterization of the spatial proteome.