Weak and Transient Protein Interactions Determined by Solid-State NMR
Weak and Transient Protein Interactions Determined by Solid-State NMR
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DOI：
10.1002/anie.201511609
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发表时间：
2016-06-01
影响因子：
16.6
通讯作者：
Pintacuda, Guido
中科院分区：
文献类型：
--
作者：
Dannatt, Hugh R. W.;Felletti, Michele;Pintacuda, Guido
尽管它们在控制许多细胞过程中发挥作用，但大结构大分子和无序蛋白质片段之间的微弱且短暂的相互作用目前无法通过 X 射线晶体学或溶液核磁共振以原子分辨率来表征。固态核磁共振不受溶液核磁共振分子尺寸限制的影响，可应用于不同聚集状态的分子，包括非晶沉淀物和沉积物。基于高磁场、快速魔角样品旋转和氘化的固态核磁共振方法提供了化学位移和弛豫图谱，能够表征 80 kDa 蛋白质组装体中两个区域之间瞬时关联的结构和动力学。这导致了对大肠杆菌 DNA 代谢调节机制的直接验证。
Despite their roles in controlling many cellular processes, weak and transient interactions between large structured macromolecules and disordered protein segments cannot currently be characterized at atomic resolution by Xray crystallography or solution NMR. Solid-state NMR does not suffer from the molecular size limitations affecting solution NMR, and it can be applied to molecules in different aggregation states, including non-crystalline precipitates and sediments. A solid-state NMR approach based on high magnetic fields, fast magic-angle sample spinning, and deuteration provides chemical-shift and relaxation mapping that enabled the characterization of the structure and dynamics of the transient association between two regions in an 80 kDa protein assembly. This led to direct verification of a mechanism of regulation of E. coli DNA metabolism.