Harden Bradley J, Frueh Dominique P
Biophysics and Biophysical Chemistry, Johns Hopkins University School of Medicine, 725 N. Wolfe Street, Baltimore, MD, 21205, USA.
Chembiochem. 2017 Apr 4;18(7):629-632. doi: 10.1002/cbic.201700030. Epub 2017 Feb 22.
Nonribosomal peptide synthetases (NRPSs) employ multiple domains separated by linker regions to incorporate substrates into natural products. During synthesis, substrates are covalently tethered to carrier proteins that translocate between catalytic partner domains. The molecular parameters that govern translocation and associated linker remodeling remain unknown. Here, we used NMR to characterize the structure, dynamics, and invisible states of a peptidyl carrier protein flanked by its linkers. We showed that the N-terminal linker stabilizes and interacts with the protein core while modulating dynamics at specific sites involved in post-translational modifications and/or domain interactions. The results detail the molecular communication between peptidyl carrier proteins and their linkers and could guide efforts in engineering NRPSs to obtain new pharmaceuticals.
非核糖体肽合成酶（NRPSs）利用由连接区隔开的多个结构域将底物整合到天然产物中。在合成过程中，底物共价连接到在催化伙伴结构域之间转运的载体蛋白上。控制转运和相关连接区重塑的分子参数仍然未知。在这里，我们使用核磁共振（NMR）来表征肽基载体蛋白及其连接区两侧的结构、动力学和不可见状态。我们表明，N端连接区在调节参与翻译后修饰和/或结构域相互作用的特定位点的动力学时，会稳定肽基载体蛋白并与其核心相互作用。这些结果详细阐述了肽基载体蛋白与其连接区之间的分子通讯，并可为改造NRPSs以获得新药物的研究提供指导。
