Anticancer Agent Research Center, Korea Research Institute of Bioscience and Biotechnology, Ochang-eup, Cheongju-si, Chungcheongbuk-do, 28116, Republic of Korea.
Department of Biomolecular Science, KRIBB School, University of Science and Technology, Daejeon, 34113, Republic of Korea.
Cell Mol Life Sci. 2021 Apr;78(7):3725-3741. doi: 10.1007/s00018-021-03805-x. Epub 2021 Mar 9.
Protein arginylation is a critical regulator of a variety of biological processes. The ability to uncover the global arginylation pattern and its associated signaling pathways would enable us to identify novel disease targets. Here, we report the development of a tool able to capture the N-terminal arginylome. This tool, termed R-catcher, is based on the ZZ domain of p62, which was previously shown to bind N-terminally arginylated proteins. Mutating the ZZ domain enhanced its binding specificity and affinity for Nt-Arg. R-catcher pulldown coupled to LC-MS/MS led to the identification of 59 known and putative arginylated proteins. Among these were a subgroup of novel ATE1-dependent arginylated ER proteins that are linked to diverse biological pathways, including cellular senescence and vesicle-mediated transport as well as diseases, such as Amyotrophic Lateral Sclerosis and Alzheimer's disease. This study presents the first molecular tool that allows the unbiased identification of arginylated proteins, thereby unlocking the arginylome and provide a new path to disease biomarker discovery.
蛋白质精氨酸化是多种生物过程的关键调节剂。能够揭示全局精氨酸化模式及其相关信号通路，将使我们能够识别新的疾病靶点。在这里，我们报告了一种能够捕获 N 端精氨酸组的工具的开发。该工具称为 R-catcher，基于先前显示与 N 端精氨酸化蛋白结合的 p62 的 ZZ 结构域。突变 ZZ 结构域增强了其对 Nt-Arg 的结合特异性和亲和力。R-catcher 下拉与 LC-MS/MS 相结合，鉴定出 59 种已知和推定的精氨酸化蛋白。其中包括一组新的 ATE1 依赖性精氨酸化 ER 蛋白，这些蛋白与多种生物途径有关，包括细胞衰老和囊泡介导的运输，以及疾病，如肌萎缩侧索硬化症和阿尔茨海默病。这项研究提出了第一个允许无偏鉴定精氨酸化蛋白的分子工具，从而揭示了精氨酸组，并为疾病生物标志物的发现提供了新途径。
