Division of Biology, California Institute of Technology, Pasadena, CA 91125, USA; Max Planck Institute for Brain Research, D-60528 Frankfurt am Main, Germany.
Curr Opin Chem Biol. 2013 Oct;17(5):738-46. doi: 10.1016/j.cbpa.2013.07.021. Epub 2013 Aug 9.
The non-canonical amino acid labeling techniques BONCAT (bioorthogonal non-canonical amino acid tagging) and FUNCAT (fluorescent non-canonical amino acid tagging) enable the specific identification and visualization of newly synthesized proteins. Recently, these techniques have been applied to neuronal systems to elucidate protein synthesis dynamics during plasticity, identify stimulation-induced proteomes and subproteomes and to investigate local protein synthesis in specific subcellular compartments. The next generation of tools and applications, reviewed here, includes the development of new tags, the quantitative identification of newly synthesized proteins, the application of NCAT to whole animals, and the ability to genetically restrict NCAT labeling. These techniques will enable not only improved detection but also allow new scientific questions to be tackled.
非经典氨基酸标记技术 BONCAT（生物正交非经典氨基酸标记）和 FUNCAT（荧光非经典氨基酸标记）可实现新合成蛋白质的特异性鉴定和可视化。最近，这些技术已被应用于神经元系统，以阐明可塑性过程中蛋白质合成的动力学，鉴定刺激诱导的蛋白质组和亚蛋白质组，并研究特定亚细胞区室中的局部蛋白质合成。本文综述了下一代工具和应用，包括新型标签的开发、新合成蛋白质的定量鉴定、NCAT 在全动物中的应用以及基因限制 NCAT 标记的能力。这些技术不仅将提高检测能力，还将能够解决新的科学问题。
