Morgan Brittany R, Zitzewitz Jill A, Massi Francesca
Department of Biochemistry and Molecular Pharmacology, University of Massachusetts, Worcester, Massachusetts.
Department of Biochemistry and Molecular Pharmacology, University of Massachusetts, Worcester, Massachusetts.
Biophys J. 2017 Aug 8;113(3):540-549. doi: 10.1016/j.bpj.2017.06.049.
Amyotrophic lateral sclerosis (ALS) is the most common adult degenerative motor neuron disease. Experimental evidence indicates a direct role of transactive-response DNA-binding protein 43 (TDP-43) in the pathology of ALS and other neurodegenerative diseases. TDP-43 has been identified as a major component of cytoplasmic inclusions in patients with sporadic ALS; however, the molecular basis of the disease mechanism is not yet fully understood. Fragmentation within the second RNA recognition motif (RRM2) of TDP-43 has been observed in patient tissues and may play a role in the formation of aggregates in disease. To determine the structural and dynamical changes resulting from the truncation that could lead to aggregation and toxicity, we performed molecular dynamics simulations of the full-length RRM2 domain (the stability core of TDP-43) and of a truncated variant (where residues 189-207 are deleted to mimic a site of cleavage within RRM2 found in ALS patients). Our simulations show heterogeneous structural reorganization and decreased stability of the truncated RRM2 domain compared to the full-length domain, consistent with previous experimental results. The decreased stability and structural reorganization in the truncated RRM2 result in a higher probability of protein-protein interactions through altered electrostatic surface charges and increased accessibility of hydrophobic residues (including the nuclear export sequence), providing a rationale for the increased cytoplasmic aggregation of RRM2 fragments seen in sporadic ALS patients.
肌萎缩侧索硬化症(ALS)是最常见的成人退行性运动神经元疾病。实验证据表明,反式作用应答DNA结合蛋白43(TDP-43)在ALS和其他神经退行性疾病的病理过程中起直接作用。TDP-43已被确定为散发性ALS患者胞质内含物的主要成分;然而,疾病机制的分子基础尚未完全阐明。在患者组织中观察到TDP-43的第二个RNA识别基序(RRM2)内的片段化,其可能在疾病中聚集体的形成中起作用。为了确定可能导致聚集和毒性的截断所引起的结构和动力学变化,我们对全长RRM2结构域(TDP-43的稳定性核心)和截短变体(其中删除了189-207位残基以模拟在ALS患者中发现的RRM2内的切割位点)进行了分子动力学模拟。我们的模拟显示,与全长结构域相比,截短的RRM2结构域存在异质性结构重组且稳定性降低,这与先前的实验结果一致。截短的RRM2中稳定性降低和结构重组导致通过改变的静电表面电荷和增加的疏水残基(包括核输出序列)可及性而发生蛋白质-蛋白质相互作用的可能性更高,这为散发性ALS患者中所见的RRM2片段胞质聚集增加提供了理论依据。