Elucidating the molecular programming of a nonlinear nonribosomal peptide synthetase responsible for fungal siderophore biosynthesis
Elucidating the molecular programming of a nonlinear nonribosomal peptide synthetase responsible for fungal siderophore biosynthesis
复制标题
阐明负责真菌铁载体生物合成的非线性非核糖体肽合成酶的分子编程
DOI：
10.1101/2022.10.10.511241
复制
发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Jenner M
中科院分区：
文献类型：
--
作者：
Jenner M
铁载体属于铁色素家族，对真菌物种的生存是必不可少的，对许多病原真菌的毒力起着关键作用。尽管它们具有生物学意义，但我们对这些铁螯合环状六肽是如何由非核糖体多肽合成酶(NRPS)组装的了解很少，主要是由于结构域结构的非线性。在这里，我们报告了SIDC NRPS的生化特征，它负责构建细胞内铁载体铁蛋白。纯化的SIDC的体外重组揭示了其产生铁蛋白及其结构变体铁色素的能力。完整蛋白质质谱仪的应用揭示了多肽铁载体生物合成过程中的几个非典型事件，包括氨基酸底物的模块间负载和能够形成聚酰胺键的腺化结构域。这项工作扩大了NRPS编程的范围，允许对铁铬NRPS进行生物合成分配，并为向新型异羟甲酸酯支架进行重新编程奠定了基础。
Siderophores belonging to the ferrichrome family are essential for the viability of fungal species and play a key role for virulence of numerous pathogenic fungi. Despite their biological significance, our understanding of how these iron-chelating cyclic hexapeptides are assembled by non-ribosomal peptide synthetase (NRPS) enzymes remains poorly understood, primarily due to the nonlinearity exhibited by the domain architecture. Herein, we report the biochemical characterization of the SidC NRPS, responsible for construction of the intracellular siderophore ferricrocin. In vitro reconstitution of purified SidC reveals its ability to produce ferricrocin and its structural variant, ferrichrome. Application of intact protein mass spectrometry uncovers several non-canonical events during peptidyl siderophore biosynthesis, including inter-modular loading of amino acid substrates and an adenylation domain capable of poly-amide bond formation. This work expands the scope of NRPS programming, allows biosynthetic assignment of ferrichrome NRPSs, and sets the stage for reprogramming towards novel hydroxamate scaffolds.