Pyrazinoic Acid Inhibits Mycobacterial Coenzyme A Biosynthesis by Binding to Aspartate Decarboxylase PanD.
Pyrazinoic Acid Inhibits Mycobacterial Coenzyme A Biosynthesis by Binding to Aspartate Decarboxylase PanD.
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DOI：
10.1021/acsinfecdis.7b00079
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发表时间：
2017-11-10
影响因子：
5.3
通讯作者：
Dick T
中科院分区：
文献类型：
--
作者：
Gopal P;Nartey W;Ragunathan P;Sarathy J;Kaya F;Yee M;Setzer C;Manimekalai MSS;Dartois V;Grüber G;Dick T
在此之前，我们发现了对吡嗪酸（pyrazinoic acid， POA）（结核病前药吡嗪酰胺（pyrazinamide， PZA）的生物活性成分）耐药的主要体外和体内机制，涉及辅酶a生物合成所需的天冬氨酸脱羧酶PanD的错义突变。POA的作用机制是什么？在证明用POA处理牛卡介苗会导致细胞内辅酶a的耗竭，并证实POA介导的耗竭可以通过PanD的错义突变或外源性泛酸补充来阻止，我们假设POA与PanD结合，这种结合阻断了生物合成途径。在这里，我们证实了这两个假设。首先，代谢组学分析表明，POA处理导致pand介导的催化步骤下游所有辅酶a前体浓度降低。其次，使用等温滴定量热法，我们确定了POA，而不是其前药PZA，与PanD结合。突变体PanD蛋白的结合被取消。综上所述，这些发现支持了POA的作用机制，其中PZA的生物活性成分通过与天冬氨酸脱羧酶PanD结合抑制辅酶a的生物合成。与之前的工作一起，这些结果确定了PanD作为PZA的遗传，代谢和生物物理验证靶标。
Previously, we showed that a major in vitro and in vivo mechanism of resistance to pyrazinoic acid (POA), the bioactive component of the critical tuberculosis (TB) prodrug pyrazinamide (PZA), involves missense mutations in the aspartate decarboxylase PanD, an enzyme required for coenzyme A biosynthesis. What is the mechanism of action of POA? Upon demonstrating that treatment of M. bovis BCG with POA resulted in a depletion of intracellular coenzyme A and confirming that this POA-mediated depletion is prevented by either missense mutations in PanD or exogenous supplementation of pantothenate, we hypothesized that POA binds to PanD and that this binding blocks the biosynthetic pathway. Here, we confirm both hypotheses. First, metabolomic analyses showed that POA treatment resulted in a reduction of the concentrations of all coenzyme A precursors downstream of the PanD-mediated catalytic step. Second, using isothermal titration calorimetry, we established that POA, but not its prodrug PZA, binds to PanD. Binding was abolished for mutant PanD proteins. Taken together, these findings support a mechanism of action of POA in which the bioactive component of PZA inhibits coenzyme A biosynthesis via binding to aspartate decarboxylase PanD. Together with previous works, these results establish PanD as a genetically, metabolically, and biophysically validated target of PZA.
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影响因子：
5.3
作者：
Irwin SM;Prideaux B;Lyon ER;Zimmerman MD;Brooks EJ;Schrupp CA;Chen C;Reichlen MJ;Asay BC;Voskuil MI;Nuermberger EL;Andries K;Lyons MA;Dartois V;Lenaerts AJ
通讯作者：
Lenaerts AJ
DOI：
10.1016/s0006-291x(02)02468-3
发表时间：
2002-11-01
影响因子：
3.1
作者：
Grüber, G;Godovac-Zimmermann, J;Bailer, SM
通讯作者：
Bailer, SM
影响因子：
6.1
作者：
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通讯作者：
Svergun, DI
影响因子：
2.9
作者：
Gopalan, Gayathri;Chopra, Sidharth;Swaminathan, Kunchithapadam
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Swaminathan, Kunchithapadam
影响因子：
5.4
作者：
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通讯作者：
Mueller, Volker