Covalent Inactivation of Mycobacterium tuberculosis Isocitrate Lyase by cis-2,3-Epoxy-Succinic Acid.
Covalent Inactivation of Mycobacterium tuberculosis Isocitrate Lyase by cis-2,3-Epoxy-Succinic Acid.
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顺式 2,3-环氧琥珀酸共价灭活结核分枝杆菌异柠檬酸裂解酶。
DOI：
10.1021/acschembio.0c00740
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发表时间：
2021
影响因子：
4
通讯作者：
Meek,ThomasD
中科院分区：
文献类型：
--
作者：
Pham,TrucViet;Mellott,DrakeM;Moghadamchargari,Zahra;Chen,Kevin;Krieger,Inna;Laganowsky,Arthur;Sacchettini,JamesC;Meek,ThomasD
异柠檬酸裂解酶（ICL 1/2）是结核分枝杆菌（Mycobacterium tuberculosis，Mtb）的必需酶。目前，没有ICL 1/2抑制剂进展到临床评价，尽管广泛的药物发现的努力。在此，我们调查了针对ICL 1的琥珀酸酯类似物，并发现限制在其同平面构象的二羧酸，如马来酸，包含ICL 1的非竞争性抑制剂，并且比其反式异构体抑制更有效。由此，我们鉴定了顺式-2，3环氧琥珀酸（cis-EpS）作为MtbICL 1的选择性、不可逆共价灭活剂（kinact/Kinact=（5.0 ± 1.4）× 104 M-1 s-1;Kinact= 200 ± 50 nM），这是迄今为止最有效的ICL 1灭活剂。晶体学和质谱分析表明，ICL 1的Cys 191被顺式-EpS S-马来酰化，并且失活的晶体学“快照”使我们深入了解这种失活的化学机制。E.大肠杆菌裂解物显示cis-EpS选择性标记质粒表达的MtbICL 1。一致的是，顺式EpS，而不是它的反式异构体，抑制Mtb的生长条件下，ICL功能是必不可少的。这些发现促进了顺式-2，3-环氧琥珀酸酯类似物作为抗结核药物的开发。
The isocitrate lyases (ICL1/2) are essential enzymes ofMycobacterium tuberculosis(Mtb), the causative agent of tuberculosis. At present, no ICL1/2 inhibitors have progressed to clinical evaluation, despite extensive drug discovery efforts. Herein, we surveyed succinate analogs against ICL1 and found that dicarboxylic acids constrained in theirsynperiplanarconformations, such as maleic acid, comprise uncompetitive inhibitors of ICL1 and inhibit more potently than theirtrans-isomers. From this, we identifiedcis-2,3 epoxysuccinic acid (cis-EpS) as a selective, irreversible covalent inactivator ofMtbICL1 (kinact/Kinact= (5.0 ± 1.4) × 104M–1s–1;Kinact= 200 ± 50 nM), the most potent inactivator of ICL1 yet characterized. Crystallographic and mass spectrometric analysis demonstrated that Cys191of ICL1 was S-malylated bycis-EpS, and a crystallographic “snapshot” of inactivation lent insight into the chemical mechanism of this inactivation. Proteomic analysis ofE. colilysates showed thatcis-EpS selectively labeled plasmid-expressedMtbICL1. Consistently,cis-EpS, but not itstrans-isomer, inhibited the growth ofMtbunder conditions in which ICL function is essential. These findings encourage the development of analogs ofcis-2,3-epoxysuccinate as antituberculosis agents.