Inhibitor Screen Identifies Covalent Inhibitors of the Protein Histidine Phosphatase PHPT1.
Inhibitor Screen Identifies Covalent Inhibitors of the Protein Histidine Phosphatase PHPT1.
复制标题
抑制剂筛选可鉴定蛋白质组氨酸磷酸酶 PHPT1 的共价抑制剂。
DOI：
10.1021/acsmedchemlett.2c00053
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发表时间：
2022
影响因子：
4.2
通讯作者：
Barrios,AmyM
中科院分区：
文献类型：
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作者：
McCullough,BrandonS;Wang,Hanfei;Barrios,AmyM
蛋白质组氨酸磷酸酶PHPT 1涉及多种细胞信号传导途径。然而，人们对这种酶的确切生物学作用知之甚少，缺乏研究组氨酸磷酸化和去磷酸化的化学工具阻碍了该领域的进展。为了确定PHPT 1活性的第一个抑制剂，我们使用我们实验室最近开发的PHPT 1活性的简易荧光测定法进行了抑制剂筛选。从一组约4000个化合物获得的微源光谱收集和NCI多样性集IV，我们确定了几个潜在的命中具有显着的选择性抑制PHPT 1活性超过其他磷酸酶。其中，去甲静酸是PHPT 1活性的最有效抑制剂，在我们的测定条件下IC 50值为7.9 ± 0.8 μM。降粘酸是PHPT 1活性的时间依赖性共价抑制剂，KI = 90 ± 20 μM，kinact = 1.7 ± 0.1 min-1。
The protein histidine phosphatase PHPT1 is implicated in a variety of cellular signaling pathways. However, little is known about the precise biological roles of this enzyme and a dearth of chemical tools for studying histidine phosphorylation and dephosphorylation has hampered progress in the field. With the goal of identifying the first inhibitors of PHPT1 activity, we carried out an inhibitor screen using a facile fluorogenic assay for PHPT1 activity recently developed in our laboratory. From a panel of approximately 4000 compounds obtained from the Microsource Spectrum Collection and the NCI Diversity Set IV, we identified several potential hits with significant selectivity for inhibiting PHPT1 activity over other phosphatases. Of these, norstictic acid was the most potent inhibitor of PHPT1 activity with an IC50value of 7.9 ± 0.8 μM under our assay conditions. Norstictic acid is a time-dependent, covalent inhibitor of PHPT1 activity withKI= 90 ± 20 μM andkinact= 1.7 ± 0.1 min–1.