Ion Mobility Mass Spectrometry Measures the Conformational Landscape of p27 and its Domains and how this is Modulated upon Interaction with Cdk2/cyclin A
Ion Mobility Mass Spectrometry Measures the Conformational Landscape of p27 and its Domains and how this is Modulated upon Interaction with Cdk2/cyclin A
复制标题
离子淌度质谱法测量 p27 及其结构域的构象景观，以及如何在与 Cdk2/cyclin A 相互作用时对其进行调节
DOI：
10.1002/ange.201812697
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发表时间：
2019
影响因子：
--
通讯作者：
Beveridge R
中科院分区：
文献类型：
--
作者：
Beveridge R
据报道，本质上无序的蛋白质在与细胞中的伙伴结合后会经历无序到有序的转变。使用经典的结构测定方法很难可视化结合时的排序程度和结合之前的缺乏顺序。 p27 与 Cdk2/cyclin A 复合物的结合伴随着 p27 在 KID 结构域中的部分折叠，保留了功能的动态行为，特别是在蛋白质的 C 末端部分。在此，采用天然离子淌度质谱（IM-MS）来测量 p27 的内在动态特性，无论是在分离状态还是在与 Cdk2/cyclin A 的三聚体复合物中。与 Cdk2/cyclin A 相比，三聚体 Cdk2/cyclin A/p27-KID 复合物具有显着的结构异质性。这些发现支持了模糊复合物的形成，其中 N 端和 C 端均具有p27 与 Cdk2/cyclin A 在多种密切相关的状态下相互作用。
Intrinsically disordered proteins have been reported to undergo disorder‐to‐order transitions upon binding to their partners in the cell. The extent of the ordering upon binding and the lack of order prior to binding is difficult to visualize with classical structure determination methods. Binding of p27 to the Cdk2/cyclin A complex is accompanied by partial folding of p27 in the KID domain, with the retention of dynamic behavior for function, particularly in the C‐terminal half of the protein. Herein, native ion mobility mass spectrometry (IM‐MS) is employed to measure the intrinsic dynamic properties of p27, both in isolation and within the trimeric complex with Cdk2/cyclin A. The trimeric Cdk2/cyclin A/p27‐KID complex possesses significant structural heterogeneity compared to Cdk2/cyclin A. These findings support the formation of a fuzzy complex in which both the N‐ and C‐termini of p27 interact with Cdk2/cyclin A in multiple, closely associated states.