Synthetic cycle of the initiation module of a formylating nonribosomal peptide synthetase
Synthetic cycle of the initiation module of a formylating nonribosomal peptide synthetase
复制标题
DOI：
10.1038/nature16503
复制
发表时间：
2016-01-14
期刊：
影响因子：
64.8
通讯作者：
Schmeing, T. Martin
中科院分区：
文献类型：
--
作者：
Reimer, Janice M.;Aloise, Martin N.;Schmeing, T. Martin
非核糖体肽合成酶（NRPS）是一种非常大的蛋白质，可产生具有广泛生物活性的小肽分子，包括环境友好的化学品和许多广泛使用的治疗药物（1）。NRPS是大分子机器，具有模块化装配线逻辑，复杂的催化循环，移动部件和许多活性位点（2，3）。除了连接底物所需的核心结构域外，它们通常还包括专门的定制结构域，这些结构域引入化学修饰并允许产品进入大片化学空间（3，4）。NRPS剪裁结构域如何在结构上被容纳到巨酶中，或者它们如何适应非核糖体肽合成中的功能，仍然是未知的。在这里，我们提出了一系列的晶体结构的起始模块的一个杀虫剂生产NRPS，线性短杆菌肽合成酶（5，6）。这个模块包括专门的裁剪领域，并在初始模块中捕获表示装配线合成的每个主要步骤的状态。构象之间的转换是大规模的，肽基载体蛋白结构域和腺苷酸化亚结构域经历巨大的运动，以运输底物之间的远端活性位点。这些结构突出了NRPS的多功能性，因为小结构域重新利用和回收它们有限的界面与它们的各种结合伴侣相互作用。如果NRPS要用于生产新的治疗剂，了解剪裁结构域是重要的。
Nonribosomal peptide synthetases (NRPSs) are very large proteins that produce small peptide molecules with wide-ranging biological activities, including environmentally friendly chemicals and many widely used therapeutics(1). NRPSs are macromolecular machines, with modular assembly-line logic, a complex catalytic cycle, moving parts and many active sites(2,3). In addition to the core domains required to link the substrates, they often include specialized tailoring domains, which introduce chemical modifications and allow the product to access a large expanse of chemical space(3,4). It is still unknown how the NRPS tailoring domains are structurally accommodated into megaenzymes or how they have adapted to function in nonribosomal peptide synthesis. Here we present a series of crystal structures of the initiation module of an antibiotic-producing NRPS, linear gramicidin synthetase(5,6). This module includes the specialized tailoring formylation domain, and states are captured that represent every major step of the assembly-line synthesis in the initiation module. The transitions between conformations are large in scale, with both the peptidyl carrier protein domain and the adenylation subdomain undergoing huge movements to transport substrate between distal active sites. The structures highlight the great versatility of NRPSs, as small domains repurpose and recycle their limited interfaces to interact with their various binding partners. Understanding tailoring domains is important if NRPSs are to be utilized in the production of novel therapeutics.