Saha Sougata, Wang Junling, Kashina Anna S
Tezpur University, Napaam, Assam, India.
University of Pennsylvania, Philadelphia, PA, USA.
Methods Mol Biol. 2023;2620:119-122. doi: 10.1007/978-1-0716-2942-0_16.
Here, we describe the biochemical assay for ATE1-mediated arginylation in microplate format, which can be applied to high-throughput screens for the identification of small molecule inhibitors and activators of ATE1, high-volume analysis of AE1 substrates, and other similar applications. Originally, we have applied this screen to a library of 3280 compounds and identified 2 compounds which specifically affect ATE1-regulated processes in vitro and in vivo. The assay is based on in vitro ATE1-mediated arginylation of beta-actin's N-terminal peptide, but it can also be applied using other ATE1 substrates.
在此，我们描述了一种微孔板形式的用于检测ATE1介导的精氨酰化的生化分析方法，该方法可应用于高通量筛选以鉴定ATE1的小分子抑制剂和激活剂、对ATE1底物进行大量分析以及其他类似应用。最初，我们已将此筛选方法应用于一个包含3280种化合物的文库，并鉴定出2种在体外和体内特异性影响ATE1调控过程的化合物。该分析基于体外ATE1介导的β-肌动蛋白N端肽的精氨酰化，但也可使用其他ATE1底物进行应用。
