Structures of a Nonribosomal Peptide Synthetase Module Bound to MbtH-like Proteins Support a Highly Dynamic Domain Architecture
Structures of a Nonribosomal Peptide Synthetase Module Bound to MbtH-like Proteins Support a Highly Dynamic Domain Architecture
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DOI：
10.1074/jbc.m116.746297
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发表时间：
2016-10-21
影响因子：
4.8
通讯作者：
Gulick, Andrew M.
中科院分区：
文献类型：
--
作者：
Miller, Bradley R.;Drake, Eric J.;Gulick, Andrew M.
非核糖体肽合成酶（NRPS）产生多种肽类天然产物。在合成过程中，多结构域NRPS充当装配线，将生长的产物从一个模块传递到下一个模块。每个模块通常由整合的肽基载体蛋白、装载氨基酸的腺苷酸化结构域和催化肽键形成的缩合结构域组成。一些腺苷酸化结构域与称为MbtH样蛋白（MLP）的小伴侣蛋白相互作用，增强溶解度或活性。结合到腺苷酸化结构域的MLP的结构先前已经使用截短的腺苷酸化结构域报道，排除了可能来自于理解MLP对完整的腺苷酸化结构域或对整个NRPS模块的动力学的影响的任何见解。在这里，我们提出了全长NRPS EntF的结构结合的MLP从大肠杆菌和铜绿假单胞菌。这些新的结构，沿着生物化学和生物信息学的支持，进一步阐述了定义MLP-腺苷酸化结构域界面的残基。此外，这些结构突出了NRPS模块的动态行为，包括由腺苷酸化和缩合结构域形成的模块核心以及移动的硫酯酶结构域的方向。
Nonribosomal peptide synthetases (NRPSs) produce a wide variety of peptide natural products. During synthesis, the multidomain NRPSs act as an assembly line, passing the growing product from one module to the next. Each module generally consists of an integrated peptidyl carrier protein, an amino acid-loading adenylation domain, and a condensation domain that catalyzes peptide bond formation. Some adenylation domains interact with small partner proteins called MbtH-like proteins (MLPs) that enhance solubility or activity. A structure of an MLP bound to an adenylation domain has been previously reported using a truncated adenylation domain, precluding any insight that might be derived from understanding the influence of the MLP on the intact adenylation domain or on the dynamics of the entire NRPS module. Here, we present the structures of the full-length NRPS EntF bound to the MLPs from Escherichia coli and Pseudomonas aeruginosa. These new structures, along with biochemical and bioinformatics support, further elaborate the residues that define the MLP-adenylation domain interface. Additionally, the structures highlight the dynamic behavior of NRPS modules, including the module core formed by the adenylation and condensation domains as well as the orientation of the mobile thioesterase domain.