Tracking carrier protein motions with Raman spectroscopy
Tracking carrier protein motions with Raman spectroscopy
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DOI：
10.1038/s41467-019-10184-2
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发表时间：
2019-05-20
影响因子：
16.6
通讯作者：
Charkoudian, Louise K.
中科院分区：
文献类型：
--
作者：
Epstein, Samuel C.;Huff, Adam R.;Charkoudian, Louise K.
工程微生物生物合成途径代表了一个引人注目的途径，以获得扩大的化学多样性。载体蛋白（CPs）在生物合成中起着核心作用，但CPs的快速运动使其构象动力学难以用传统的光谱方法捕获。在这里，我们提出了一种低资源的方法来直接揭示载体蛋白与底物的相互作用。化学酶将市售的含炔的底物装载到CPs上，可以使用拉曼光谱快速可视化分子货物的局部环境。该方法可以明确链固存机制的基础，促进CPs的快速表征，并使天然产物在体外和体内的载体加工可视化。
Engineering microbial biosynthetic pathways represents a compelling route to gain access to expanded chemical diversity. Carrier proteins (CPs) play a central role in biosynthesis, but the fast motions of CPs make their conformational dynamics difficult to capture using traditional spectroscopic approaches. Here we present a low-resource method to directly reveal carrier protein-substrate interactions. Chemoenzymatic loading of commercially available, alkyne-containing substrates onto CPs enables rapid visualization of the molecular cargo's local environment using Raman spectroscopy. This method could clarify the foundations of the chain sequestration mechanism, facilitate the rapid characterization of CPs, and enable visualization of the vectoral processing of natural products both in vitro and in vivo.