Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Katsura, Nishikyo-ku, Kyoto 615-8510, Japan.
Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Katsura, Nishikyo-ku, Kyoto 615-8510, Japan; Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency, Sanbancho, Chiyodaku, Tokyo 102-0075, Japan.
Curr Opin Chem Biol. 2019 Feb;48:1-7. doi: 10.1016/j.cbpa.2018.08.001. Epub 2018 Aug 28.
Protein functions are tightly regulated by their subcellular localization and dynamic alteration. Chemical proteomics offers convenience and efficiency for profiling protein features in a native context. In this review, we summarize the recent progress of subcellular-compartment-focused chemical proteomics which do not rely on organelle fractionation. Organelle-specific activity-based protein profiling (ABPP) and engineered ascorbate peroxidase (APEX) have been developed for proteome analysis within organelles and even sub-organelles. In parallel, our lab designed organelle-localizable reactive molecules (ORMs) to selectively tag nuclear and mitochondrial proteins. ORMs-based proteomics is applicable to primary neurons and brain slices, as well as cultured cell lines. In addition, we invented a conditional proteomics approach to elucidate zinc homeostasis by labeling and identifying proteins localized in Zn-rich space of live cells.
蛋白质的功能受到其亚细胞定位和动态变化的严格调控。化学蛋白质组学为在天然环境中分析蛋白质特征提供了便利和效率。在这篇综述中，我们总结了近年来非基于细胞器分离的亚细胞区室聚焦化学蛋白质组学的进展。细胞器特异性基于活性的蛋白质谱分析（ABPP）和工程化抗坏血酸过氧化物酶（APEX）已被开发用于细胞器甚至亚细胞器中的蛋白质组分析。同时，我们实验室设计了细胞器定位反应性分子（ORMs），以选择性标记核和线粒体蛋白。基于 ORMs 的蛋白质组学适用于原代神经元和脑切片，以及培养的细胞系。此外，我们发明了一种条件蛋白质组学方法，通过标记和鉴定活细胞中富含锌的空间定位的蛋白质来阐明锌稳态。
