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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DOI：
10.1002/anie.202014457
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发表时间：
2021-06-07
期刊：
影响因子：
--
通讯作者：
Tate EW
中科院分区：
文献类型：
--
作者：
Lanyon-Hogg T;Ritzefeld M;Zhang L;Andrei SA;Pogranyi B;Mondal M;Sefer L;Johnston CD;Coupland CE;Greenfield JL;Newington J;Fuchter MJ;Magee AI;Siebold C;Tate EW
哺乳动物膜结合O-酰基转移酶(MBOAT)超家族参与生长、发育和食欲感觉等生物学过程。MBOAT是治疗癌症和肥胖症的有吸引力的药物靶点；然而，关于小分子抑制作用的结合部位和分子机制的信息尚不清楚。这项研究报告了一种合理的光化学探针，用于询问人MBOAT Hedgehog酰基转移酶(HHAT)中一个新的小分子抑制物结合位点。构效关系研究确定了迄今为止报道的最有效的HHAT抑制剂IMP-1575的单一对映体，并指导设计了保持HHAT抑制效力的光交联探针。HHAT的光交联和蛋白质组学测序提供了哺乳动物MBOAT中第一个小分子结合位点的鉴定。拓扑和同源性数据提示了HHAT抑制的潜在机制，动力学分析证实了这一点。我们的结果提供了一个最佳的HHAT工具抑制剂IMP-1575(Ki=38 nm)，并提供了一种在MBOAT中定位小分子相互作用位点的策略。Hedgehog酰基转移酶(HHAT)抑制剂的构效关系分析有助于合理设计HHAT的光化学探针。探针光交联法发现了HHAT中第一个小分子抑制剂结合部位，揭示了HHAT的抑制机制，为进一步研究提供了最佳的HHAT工具抑制剂IMP-1575(Ki=38 nM)。
The mammalian membrane‐bound O‐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 enantiomer IMP‐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 inhibitor IMP‐1575 (K i=38 nM) and a strategy for mapping small molecule interaction sites in MBOATs. Structure‐activity relationship analysis of Hedgehog acyltransferase (HHAT) inhibitors allowed rational design of photochemical probes for HHAT. Probe photocrosslinking identified the first small‐molecule inhibitor binding site in HHAT and revealed the inhibitory mechanism, providing an optimal HHAT tool inhibitor IMP‐1575 (K i=38 nM) for future studies.
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DOI：
10.1074/jbc.m114.614578
发表时间：
2015-02-06
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Konitsiotis AD;Jovanović B;Ciepla P;Spitaler M;Lanyon-Hogg T;Tate EW;Magee AI
通讯作者：
Magee AI
影响因子：
7.4
作者：
Whitelegge JP
通讯作者：
Whitelegge JP
影响因子：
15
作者：
Shieh P;Dien VT;Beahm BJ;Castellano JM;Wyss-Coray T;Bertozzi CR
通讯作者：
Bertozzi CR
影响因子：
4.2
作者：
Smith E;Collins I
通讯作者：
Collins I
影响因子：
64.5
作者：
Parker CG;Galmozzi A;Wang Y;Correia BE;Sasaki K;Joslyn CM;Kim AS;Cavallaro CL;Lawrence RM;Johnson SR;Narvaiza I;Saez E;Cravatt BF
通讯作者：
Cravatt BF