PDF(Pubmed)
Abstract：
In the quest for new bioactive substances, nonribosomal peptide synthetases (NRPS) provide biodiversity by synthesizing nonproteinaceous peptides with high cellular activity. NRPS machinery consists of multiple modules, each catalyzing a unique series of chemical reactions. Incomplete understanding of the biophysical principles orchestrating these reaction arrays limits the exploitation of NRPSs in synthetic biology. Here, we use nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry to solve the conundrum of how intermodular recognition is coupled with loaded carrier protein specificity in the tomaymycin NRPS. We discover an adaptor domain that directly recruits the loaded carrier protein from the initiation module to the elongation module and reveal its mechanism of action. The adaptor domain of the type found here has specificity rules that could potentially be exploited in the design of engineered NRPS machinery.
摘要：
为了寻找新的生物活性物质，非核糖体肽合成酶（NRPS）通过合成具有高细胞活性的非蛋白质肽来提供生物多样性。NRPS机械由多个模块组成，每个催化一系列独特的化学反应。对编排这些反应阵列的生物物理原理的不完全理解限制了NRPS在合成生物学中的利用。这里,我们使用核磁共振（NMR）光谱和质谱来解决模间识别如何与托马霉素NRPS中的负载载体蛋白特异性耦合的难题。我们发现了一个衔接子结构域，它直接将负载的载体蛋白从起始模块招募到延伸模块，并揭示了其作用机制。此处发现的类型的衔接子结构域具有特异性规则，可以在工程NRPS机器的设计中潜在地利用。
