1Department of Pharmaceutical and Biomedical Sciences, University of Georgia, Athens, GA 30602 USA.
2Present Address: The Chemical Proteomics Center and State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 201203 China.
Signal Transduct Target Ther. 2018 Mar 9;3:6. doi: 10.1038/s41392-018-0009-6. eCollection 2018.
Protein arginine methyltransferases (PRMTs) are crucial epigenetic regulators in eukaryotic organisms that serve as histone writers for chromatin remodeling. PRMTs also methylate a variety of non-histone protein substrates to modulate their function and activity. The development of potent PRMT inhibitors has become an emerging and imperative research area in the drug discovery field to provide novel therapeutic agents for treating diseases and as tools to investigate the biological functions of PRMTs. PRMT1 is the major type I enzyme that catalyzes the formation of asymmetric dimethyl arginine, and PRMT1 plays important regulatory roles in signal transduction, transcriptional activation, RNA splicing, and DNA repair. Aberrant expression of PRMT1 is found in many types of cancers, pulmonary diseases, cardiovascular disease, diabetes, and renal diseases. PRMT1 is a highly promising target for therapeutic development. We created a stopped flow fluorescence-based assay for PRMT1 inhibitor detection and characterization that has the advantages of being homogeneous, nonradioactive, and mix-and-measure in nature, allowing for continuous measurement of the methylation reaction and its inhibition. To our knowledge, this is the first continuous assay for PRMT1 reaction detection and inhibitor characterization. The approach is not only capable of quantitatively determining the potency (IC) of PRMT1 inhibitors but can also distinguish cofactor-competitive inhibitors, substrate-competitive inhibitors, and mixed-type inhibitors.
蛋白质精氨酸甲基转移酶（PRMTs）是真核生物中重要的表观遗传调控因子，作为组蛋白修饰酶参与染色质重塑。PRMTs 还可以甲基化多种非组蛋白蛋白底物，以调节其功能和活性。开发有效的 PRMT 抑制剂已成为药物发现领域的一个新兴且紧迫的研究领域，旨在为治疗疾病提供新的治疗剂，并作为研究 PRMTs 生物学功能的工具。PRMT1 是催化形成非对称二甲基精氨酸的主要 I 型酶，PRMT1 在信号转导、转录激活、RNA 剪接和 DNA 修复中发挥重要的调节作用。PRMT1 的异常表达存在于许多类型的癌症、肺部疾病、心血管疾病、糖尿病和肾脏疾病中。PRMT1 是治疗开发的一个极具前景的靶点。我们创建了一种基于停流荧光的 PRMT1 抑制剂检测和表征的方法，该方法具有均相、非放射性和混合测量的优点，允许连续测量甲基化反应及其抑制。据我们所知，这是第一个用于 PRMT1 反应检测和抑制剂表征的连续测定法。该方法不仅能够定量测定 PRMT1 抑制剂的效力（IC），还能够区分辅因子竞争性抑制剂、底物竞争性抑制剂和混合型抑制剂。
