Protein-protein interactions within the Fatty Acid Synthase-II system of Mycobacterium tuberculosis are essential for mycobacterial viability
Protein-protein interactions within the Fatty Acid Synthase-II system of Mycobacterium tuberculosis are essential for mycobacterial viability
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DOI：
10.1111/j.1365-2958.2004.04334.x
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发表时间：
2004-12-01
影响因子：
3.6
通讯作者：
Zerbib, D
中科院分区：
文献类型：
--
作者：
Veyron-Churlet, R;Guerrini, O;Zerbib, D
尽管存在有效的化疗，结核病仍然是全世界死亡的主要原因。迫切需要新药来减少病原体结核分枝杆菌 (Mtb) 多重耐药菌株出现的潜在影响。一线抗生素异烟肼 (INH) 和其他几种药物以分枝菌酸的生物合成为目标，尤其是脂肪酸合成酶-II (FAS-II) 延伸系统。这种生物合成途径对于分枝杆菌来说是必需的和特异的，并且仍然代表着寻找新的抗结核药物的有价值的系统。文献中的一些数据表明 FAS-II 系统内存在蛋白质-蛋白质相互作用。这些相互作用本身可能成为针对结核分枝杆菌的新一代药物的靶点。通过使用广泛的体内酵母双杂交方法和体外免疫共沉淀，我们证明了 FAS-II 已知成分之间存在同型和异型相互作用。缩合酶 KasA、KasB 和 mtFabH 彼此相互作用，并与还原酶 MabA 和 InhA 相互作用。此外，我们设计并构建了 FAS-II 还原酶 MabA 的点突变，能够破坏其同型相互作用并扰乱该蛋白质在 FAS-II 内的相互作用模式。最后，我们通过跨显性遗传方法表明，这些突变体在非致病性和致病性分枝杆菌中均呈显性失活。这些数据使我们能够绘制 FAS-II 组织的动态模型。它们还代表了设计新一代抗结核药物（作为必需蛋白质-蛋白质相互作用的抑制剂）的重要一步。
Despite the existence of efficient chemotherapy, tuberculosis remains a leading cause of mortality worldwide. New drugs are urgently needed to reduce the potential impact of the emergence of multidrug-resistant strains of the causative agent Mycobacterium tuberculosis (Mtb). The front-line antibiotic isoniazid (INH), and several other drugs, target the biosynthesis of mycolic acids and especially the Fatty Acid Synthase-II (FAS-II) elongation system. This biosynthetic pathway is essential and specific for mycobacteria and still represents a valuable system for the search of new anti-tuberculous agents. Several data, in the literature, suggest the existence of protein-protein interactions within the FAS-II system. These interactions themselves might serve as targets for a new generation of drugs directed against Mtb. By using an extensive in vivo yeast two-hybrid approach and in vitro co-immunoprecipitation, we have demonstrated the existence of both homotypic and heterotypic interactions between the known components of FAS-II. The condensing enzymes KasA, KasB and mtFabH interact with each other and with the reductases MabA and InhA. Furthermore, we have designed and constructed point mutations of the FAS-II reductase MabA, able to disrupt its homotypic interactions and perturb the interaction pattern of this protein within FAS-II. Finally, we showed by a transdominant genetic approach that these mutants are dominant negative in both non-pathogenic and pathogenic mycobacteria. These data allowed us to draw a dynamic model of the organization of FAS-II. They also represent an important step towards the design of a new generation of anti-tuberculous agents, as being inhibitors of essential protein-protein interactions.