Department of Chemistry, The Scripps Research Institute, Jupiter, Florida 33458, United States.
ACS Chem Biol. 2011 Oct 21;6(10):1127-35. doi: 10.1021/cb2001473. Epub 2011 Aug 23.
The protein arginine methyltransferases (PRMTs) are SAM-dependent enzymes that catalyze the mono- and dimethylation of peptidyl arginine residues. PRMT1 is the founding member of the PRMT family, and this isozyme is responsible for methylating ∼85% of the arginine residues in mammalian cells. Additionally, PRMT1 activity is aberrantly upregulated in heart disease and cancer. As a part of a program to develop isozyme-specific PRMT inhibitors, we recently described the design and synthesis of C21, a chloroacetamidine bearing histone H4 tail analogue that acts as an irreversible PRMT1 inhibitor. Given the covalent nature of the interaction, we set out to develop activity-based probes (ABPs) that could be used to characterize the physiological roles of PRMT1. Herein, we report the design, synthesis, and characterization of fluorescein-conjugated C21 (F-C21) and biotin-conjugated C21 (B-C21) as PRMT1-specific ABPs. Additionally, we provide the first evidence that PRMT1 activity is negatively regulated in a spatial and temporal fashion.
精氨酸甲基转移酶（PRMTs）是一类依赖 SAM 的酶，能够催化肽基精氨酸残基的单甲基化和二甲基化。PRMT1 是 PRMT 家族的创始成员，该同工酶负责甲基化哺乳动物细胞中约 85%的精氨酸残基。此外，PRMT1 的活性在心脏病和癌症中异常上调。作为开发同工酶特异性 PRMT 抑制剂计划的一部分，我们最近描述了 C21 的设计和合成，C21 是一种带有组蛋白 H4 尾部类似物的氯乙酰胺，可作为不可逆的 PRMT1 抑制剂。鉴于相互作用的共价性质，我们着手开发可用于表征 PRMT1 生理作用的活性探针（ABPs）。本文报告了荧光素缀合的 C21（F-C21）和生物素缀合的 C21（B-C21）作为 PRMT1 特异性 ABPs 的设计、合成和表征。此外，我们首次提供了证据表明 PRMT1 的活性受到时空调节。
