Department of Chemistry and Biochemistry, University of Delaware, Newark, Delaware 19716, USA.
Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH 43210, USA.
Sci Rep. 2017 Mar 17;7:44506. doi: 10.1038/srep44506.
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 (P) 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.
细胞肌动蛋白动力学是许多细胞过程的基本要素，例如细胞运动、细胞分裂和胞吞作用。肌动蛋白在这些过程中的参与是由许多肌动蛋白结合蛋白介导的，其中丝切蛋白家族在依赖核苷酸状态的方式中以独特和必需的方式加速丝状肌动蛋白（F-肌动蛋白）中的肌动蛋白踩踏。丝切蛋白通过识别与 ATP 水解和无机磷（P）从肌动蛋白核苷酸裂缝释放相关的 F-肌动蛋白结构的时变，优先与较旧的纤维相互作用。丝切蛋白在 F-肌动蛋白上的结构和分子间界面的细节在原子分辨率下仍知之甚少。在这里，我们通过魔角旋转（MAS）NMR 报告了肌球蛋白同工型人丝切蛋白 2（CFL2）与 F-肌动蛋白结合的原子水平特征。我们证明了大多数原子的共振分配很容易完成，并且我们推导出 CFL2 和 F-肌动蛋白之间的分子间界面。这里报道的 MAS NMR 方法为与 F-肌动蛋白结合的广泛肌动蛋白相关蛋白的原子分辨率特性建立了基础。
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