Structural Analysis of Human Cofilin 2/Filamentous Actin Assemblies: Atomic-Resolution Insights from Magic Angle Spinning NMR Spectroscopy.
Structural Analysis of Human Cofilin 2/Filamentous Actin Assemblies: Atomic-Resolution Insights from Magic Angle Spinning NMR Spectroscopy.
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DOI：
10.1038/srep44506
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发表时间：
2017-03-17
影响因子：
4.6
通讯作者：
Polenova T
中科院分区：
文献类型：
--
作者：
Yehl J;Kudryashova E;Reisler E;Kudryashov D;Polenova T
细胞肌动蛋白动力学是许多细胞过程的重要组成部分，例如细胞运动、细胞分裂和内吞作用。肌动蛋白参与这些过程是由许多肌动蛋白结合蛋白介导的，其中丝切蛋白家族在以核苷酸状态依赖性方式加速丝状肌动蛋白（F-肌动蛋白）中的肌动蛋白跑步方面发挥着独特且重要的作用。 Cofilin 通过识别与 ATP 水解和肌动蛋白核苷酸裂口释放无机磷酸盐 (Pi) 相关的 F-肌动蛋白结构的时间依赖性变化，优先与较老的肌丝相互作用。 F-肌动蛋白上肌动蛋白丝切蛋白的结构和分子间界面的细节在原子分辨率上仍然知之甚少。在此，我们报告了通过魔角旋转 (MAS) NMR 对与 F-肌动蛋白结合的人肌动蛋白丝切蛋白 2 (CFL2) 的肌肉亚型进行的原子级表征。我们证明大多数原子的共振分配很容易完成，并且我们推导了 CFL2 和 F-肌动蛋白之间的分子间界面。这里报道的 MAS NMR 方法为广泛的与 F-肌动蛋白结合的肌动蛋白相关蛋白的原子分辨率表征奠定了基础。
Cellular actin dynamics is an essential element of numerous cellular processes, such as cell motility, cell division and endocytosis. Actin’s involvement in these processes is mediated by many actin-binding proteins, among which the cofilin family plays unique and essential role in accelerating actin treadmilling in filamentous actin (F-actin) in a nucleotide-state dependent manner. Cofilin preferentially interacts with older filaments by recognizing time-dependent changes in F-actin structure associated with the hydrolysis of ATP and release of inorganic phosphate (Pi) from the nucleotide cleft of actin. The structure of cofilin on F-actin and the details of the intermolecular interface remain poorly understood at atomic resolution. Here we report atomic-level characterization by magic angle spinning (MAS) NMR of the muscle isoform of human cofilin 2 (CFL2) bound to F-actin. We demonstrate that resonance assignments for the majority of atoms are readily accomplished and we derive the intermolecular interface between CFL2 and F-actin. The MAS NMR approach reported here establishes the foundation for atomic-resolution characterization of a broad range of actin-associated proteins bound to F-actin.