Top Runner Incubation Center for Academia-Industry Fusion, Nagaoka University of Technology, 1603-1 Kamitomioka, Nagaoka, Niigata 940-2188, Japan; Department of Bioengineering, Nagaoka University of Technology, 1603-1 Kamitomioka, Nagaoka, Niigata 940-2188, Japan.
Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Katsura, Kyoto 615-8510, Japan; Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency, 5 Sanbancho, Chiyoda-ku, Tokyo 102-0075, Japan.
Curr Opin Chem Biol. 2014 Aug;21:136-43. doi: 10.1016/j.cbpa.2014.07.012. Epub 2014 Aug 13.
The ability to introduce any chemical probe to any endogenous target protein in its native environment, that is in cells and in vivo, is anticipated to provide various new exciting tools for biological and biomedical research. Although still at the prototype stage, the ligand-directed tosyl (LDT) chemistry is a novel type of affinity labeling technique that we developed for such a dream. This chemistry allows for modifying native proteins by various chemical probes with high specificity in various biological settings ranging from in vitro (in test tubes) to in living cells and in vivo. Since the first report, the list of proteins that are successfully labeled by the LDT chemistry has been increasing. A growing number of studies have demonstrated its utility to create semisynthetic proteins directly in cellular contexts. The in situ generated semisynthetic proteins are applicable for various types of analysis and imaging of intracellular biological processes. In this review, we summarize the basic properties of the LDT chemistry and its applications toward in situ engineering and analysis of native proteins in living systems. Current limitations and future challenges of this area are also described.
预期能够将任何化学探针引入任何内源性靶蛋白的天然环境中，即在细胞内和体内，这将为生物和生物医学研究提供各种令人兴奋的新工具。尽管仍处于原型阶段，但配体定向甲苯磺酰基（LDT）化学是我们为实现这一梦想而开发的一种新型亲和标记技术。这种化学可以在各种生物环境中（从试管中的体外到活细胞内和体内），用各种化学探针高度特异性地修饰天然蛋白质。自首次报道以来，成功用 LDT 化学标记的蛋白质的列表一直在增加。越来越多的研究表明，它可用于在细胞环境中直接创建半合成蛋白质。原位生成的半合成蛋白质适用于各种类型的细胞内生物过程的分析和成像。在这篇综述中，我们总结了 LDT 化学的基本性质及其在原位工程和活体系统中天然蛋白质分析中的应用。还描述了该领域当前的局限性和未来的挑战。
