Mycobacterium tuberculosis PanD Structure-Function Analysis and Identification of a Potent Pyrazinoic Acid-Derived Enzyme Inhibitor.
Mycobacterium tuberculosis PanD Structure-Function Analysis and Identification of a Potent Pyrazinoic Acid-Derived Enzyme Inhibitor.
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DOI：
10.1021/acschembio.1c00131
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发表时间：
2021-06-18
影响因子：
4
通讯作者：
Grüber G
中科院分区：
文献类型：
--
作者：
Ragunathan P;Cole M;Latka C;Aragaw WW;Hegde P;Shin J;Subramanian Manimekalai MS;Rishikesan S;Aldrich CC;Dick T;Grüber G
在抗菌药物发现中采用的一种常见策略是靶向病原体所必需的和特异性的生物合成过程。特异性特别避免了与人类宿主中潜在酶对应物的不良相互作用，并确保了靶向毒性。泛酸（维生素B5）的合成，酰基载体辅酶A的前体，是这种途径的一个例子。在结核分枝杆菌（Mycobacterium tuberculosis，Mtb）中，泛酸是由泛酸合成酶利用D-泛酸和β-Ala作为底物形成的。β-Ala主要由L-天冬氨酸脱羧形成，脱羧酶PanD在溶液中产生均低聚物。吡嗪酸（POA），TB前药吡嗪酰胺的生物活性形式，结合并抑制PanD活性弱。在这里，我们根据结构信息和PZA/POA抗性突变体产生了重组Mtb PanD突变体文库。在相应的突变体中观察到寡聚体形成，酶活性和/或POA结合的改变，为Mtb PanD的适当结构组装，脱羧活性和药物相互作用提供了必需氨基酸的见解。这些信息为设计在吡嗪环的3位具有修饰的新型POA类似物提供了平台。类似物2，在该位置引入庞大的萘酰胺基，与POA相比，显示出1,000倍的酶抑制增加，沿着适度改善的抗分枝杆菌活性。这些数据表明，对关键代谢酶的机制和酶特征的更好理解可以刺激更有效的PanD抑制剂的设计。
A common strategy employed in antibacterial drug discovery is targeting of biosynthetic processes which are essential and specific for the pathogen. Specificity in particular avoids undesirable interactions with potential enzymatic counterparts in the human host, and ensures on-target toxicity. Synthesis of pantothenate (Vitamine B5), a precursor of the acyl carrier coenzyme A, is an example of such a pathway. In Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis (TB), pantothenate is formed by pantothenate synthase, utilizing D-pantoate and β-Ala as substrates. β-Ala is mainly formed by the decarboxylation of L-aspartate, generated by the decarboxylase PanD, a homo-oliogomer in solution. Pyrazinoic acid (POA), the bioactive form of the TB prodrug pyrazinamide, binds and inhibits PanD activity weakly. Here, we generated a library of recombinant Mtb PanD mutants based on structural information and PZA/POA resistance mutants. Alterations in oligomer formation, enzyme activity and/or POA binding were observed in respective mutants, providing insights into essential amino acids for Mtb PanD’s proper structural assembly, decarboxylation activity and drug interaction. This information provided the platform for the design of novel POA analogs with modifications at position 3 of the pyrazine ring. Analog 2, incorporating a bulky naphthamido group at this position, displayed a 1,000-fold increase in enzyme inhibition compared to POA, along with moderately improved antimycobacterial activity. The data demonstrate that an improved understanding of mechanistic and enzymatic features of key metabolic enzymes can stimulate design of more potent PanD inhibitors.
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影响因子：
4.9
作者：
Lu, Ping;Haagsma, Anna C.;Bald, Dirk
通讯作者：
Bald, Dirk
影响因子：
5.3
作者：
Gopal P;Nartey W;Ragunathan P;Sarathy J;Kaya F;Yee M;Setzer C;Manimekalai MSS;Dartois V;Grüber G;Dick T
通讯作者：
Dick T
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：
5.4
作者：
Hotra, Adam;Suter, Manuel;Grueber, Gerhard
通讯作者：
Grueber, Gerhard
影响因子：
2.8
作者：
Gengenbacher, Martin;Rao, Srinivasa P. S.;Dick, Thomas
通讯作者：
Dick, Thomas