Covalent Enzyme Inhibition through Fluorosulfate Modification of a Noncatalytic Serine Residue
Covalent Enzyme Inhibition through Fluorosulfate Modification of a Noncatalytic Serine Residue
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DOI：
10.1021/acschembio.7b00403
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发表时间：
2017-08-01
影响因子：
4
通讯作者：
Jones, Lyn H.
中科院分区：
文献类型：
--
作者：
Fadeyi, Olugbeminiyi O.;Hoth, Lise R.;Jones, Lyn H.
不可逆酶抑制剂和共价化学生物探针通常利用蛋白质半胱氨酸残基与配体模板上适当定位的电泳体(例如，丙烯酰胺)的反应。然而，半胱氨酸残基并不总是可用于特定部位的蛋白质标记，因此需要新的方法来扩大以替代氨基酸为靶标的适当亲电体(“弹头”)的工具包。我们之前描述了使用带有磺酰氟电泳层的抑制剂，合理地靶向蛋白质活性部位的酪氨酸残基(信使核糖核糖核酸酶，DCPS)。这些抑制剂随后使可点击探针技术的发展成为可能，以测量活细胞中药物靶标的占有率。在这里，我们描述了一种含有氟硫酸盐的抑制剂(芳基氟硫酸盐探针(FS-p1))，它具有良好的化学和代谢稳定性，可以选择性地与DCPS中相同活性部位的非催化丝氨酸残基反应，这一点被肽图实验证实。我们的结果表明，利用氟代硫酸盐亲电弹头的非催化丝氨酸靶向可能是开发共价抑制剂药物和化学探针的合适策略。
Irreversible enzyme inhibitors and covalent chemical biology probes often utilize the reaction of a protein cysteine residue with an appropriately positioned electrophile (e.g., acrylamide) on the ligand template. However, cysteine residues are not always available for site-specific protein labeling, and therefore new approaches are needed to expand the toolkit of appropriate electrophiles ("warheads") that target alternative amino acids. We previously described the rational targeting of tyrosine residues in the active site of a protein (the mRNA decapping scavenger enzyme, DcpS) using inhibitors armed with a sulfonyl fluoride electrophile. These inhibitors subsequently enabled the development of clickable probe technology to measure drug-target occupancy in live cells. Here we describe a fluorosulfate-containing inhibitor (aryl fluorosulfate probe (FS-p1)) with excellent chemical and metabolic stability that reacts selectively with a noncatalytic serine residue in the same active site of DcpS as confirmed by peptide mapping experiments. Our results suggest that noncatalytic serine targeting using fluorosulfate electrophilic warheads could be a suitable strategy for the development of covalent inhibitor drugs and chemical probes.