Photochemical Probe Identification of a Small-Molecule Inhibitor Binding Site in Hedgehog Acyltransferase (HHAT)**
Photochemical Probe Identification of a Small-Molecule Inhibitor Binding Site in Hedgehog Acyltransferase (HHAT)**
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Hedgehog 酰基转移酶 (HHAT) 中小分子抑制剂结合位点的光化学探针鉴定**
DOI：
10.1002/ange.202014457
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发表时间：
2021
影响因子：
--
通讯作者：
Lanyon-Hogg T
中科院分区：
文献类型：
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作者：
Lanyon-Hogg T
哺乳动物膜结合O-酰基转移酶（MBOAT）超家族参与包括生长、发育和食欲感知的生物过程。MBOAT是癌症和肥胖症中有吸引力的药物靶标;然而，关于小分子抑制的结合位点和分子机制的信息是难以捉摸的。本研究报告了光化学探针的合理开发，以询问人MBOAT Hedgehog酰基转移酶（HHAT）中的新型小分子抑制剂结合位点。结构-活性关系研究确定了单一对映体IMP-1575，这是迄今为止报道的最有效的HHAT抑制剂，并指导了维持HHAT抑制效力的光交联探针的设计。HHAT的光交联和蛋白质组测序鉴定了哺乳动物MBOAT中的第一个小分子结合位点。拓扑结构和同源性数据表明HHAT抑制的潜在机制，动力学分析证实。我们的研究结果提供了一个最佳的HHAT工具包IMP-1575（Ki=38 nM）和一个在MBOAT中定位小分子相互作用位点的策略。
The mammalian membrane‐boundO‐acyltransferase (MBOAT) superfamily is involved in biological processes including growth, development and appetite sensing. MBOATs are attractive drug targets in cancer and obesity; however, information on the binding site and molecular mechanisms underlying small‐molecule inhibition is elusive. This study reports rational development of a photochemical probe to interrogate a novel small‐molecule inhibitor binding site in the human MBOAT Hedgehog acyltransferase (HHAT). Structure‐activity relationship investigation identified single enantiomerIMP‐1575, the most potent HHAT inhibitor reported to‐date, and guided design of photocrosslinking probes that maintained HHAT‐inhibitory potency. Photocrosslinking and proteomic sequencing of HHAT delivered identification of the first small‐molecule binding site in a mammalian MBOAT. Topology and homology data suggested a potential mechanism for HHAT inhibition which was confirmed by kinetic analysis. Our results provide an optimal HHAT tool inhibitorIMP‐1575(Ki=38 nM) and a strategy for mapping small molecule interaction sites in MBOATs.