The Coenzyme A Level Modulator Hopantenate (HoPan) Inhibits Phosphopantotenoylcysteine Synthetase Activity.
The Coenzyme A Level Modulator Hopantenate (HoPan) Inhibits Phosphopantotenoylcysteine Synthetase Activity.
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DOI：
10.1021/acschembio.1c00535
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发表时间：
2021-11-19
影响因子：
4
通讯作者：
Strauss E
中科院分区：
文献类型：
--
作者：
Mostert KJ;Sharma N;van der Zwaag M;Staats R;Koekemoer L;Anand R;Sibon OCM;Strauss E
泛酸类似物何首乌在细胞生物学和疾病模型中被广泛用作辅酶A(CoA)水平的调节剂，特别是在泛酸激酶相关神经退行性变(PKAN)中，这是一种源于CoA代谢受损的遗传病。HOPAN的这种使用是基于有报道称它抑制了辅酶A生物合成的第一种酶-泛酸激酶(Pank)。利用体外酶动力学研究、晶体结构分析和典型的PKAN细胞生物学模型实验相结合的方法，我们证明了HoPAN依赖于它来进行代谢激活，而不是抑制Pank。一旦被磷酸化，HoPAN通过形成一个非生产性底物复合体来抑制辅酶A途径中的下一个酶-磷酸泛硫酰半胱氨酸合成酶(PPCS)。此外，获得的人PPCS与抑制剂和激活核苷酸类似物的复合体的结构为PPCS酶的催化机制提供了新的见解-包括难以捉摸的半胱氨酸结合模式-并揭示了与严重扩张型心肌病有关的人PPCS突变的功能含义。综上所述，这项研究表明HOPAN的分子作用机制比之前认为的更复杂，这表明将其用作工具化合物的研究结果必须谨慎解释。此外，我们的发现为评估CoA导向的抑制剂效力的各种因素提供了一个明确的框架，这将被证明在未来合理开发人类遗传和传染病的潜在治疗方法中是有用的。
The pantothenate analogue hopantenate (HoPan) is widely used as a modulator of coenzyme A (CoA) levels in cell biology and disease models—especially for pantothenate kinase associated neurodegeneration (PKAN), a genetic disease rooted in impaired CoA metabolism. This use of HoPan was based on reports that it inhibits pantothenate kinase (PanK), the first enzyme of CoA biosynthesis. Using a combination of in vitro enzyme kinetic studies, crystal structure analysis, and experiments in a typical PKAN cell biology model, we demonstrate that instead of inhibiting PanK, HoPan relies on it for metabolic activation. Once phosphorylated, HoPan inhibits the next enzyme in the CoA pathway—phosphopantothenoylcysteine synthetase (PPCS)—through formation of a non-productive substrate complex. Moreover, the obtained structure of the human PPCS in complex with the inhibitor and activating nucleotide analogue provides new insights into the catalytic mechanism of PPCS enzymes—including the elusive binding mode for cysteine—and reveals the functional implications of mutations in the human PPCS that have been linked to severe dilated cardiomyopathy. Taken together, this study demonstrates that the molecular mechanism of action of HoPan is more complex than previously thought, suggesting that the results of studies in which it is used as tool compound must be interpreted with care. Moreover, our findings provide a clear framework for evaluating the various factors that contribute to the potency of CoA-directed inhibitors, one that will prove useful in the future rational development of potential therapies of both human genetic and infectious diseases.
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影响因子：
5.7
作者：
Manoj, N;Strauss, E;Ealick, SE
通讯作者：
Ealick, SE
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
影响因子：
3.2
作者：
Balibar, Carl J.;Hollis-Symynkywicz, Micah F.;Tao, Jianshi
通讯作者：
Tao, Jianshi
影响因子：
3.9
作者：
Hayflick, Susan J.
通讯作者：
Hayflick, Susan J.
影响因子：
3
作者：
Morris, Garrett M.;Huey, Ruth;Lindstrom, William;Sanner, Michel F.;Belew, Richard K.;Goodsell, David S.;Olson, Arthur J.
通讯作者：
Olson, Arthur J.