Biochemical and Biophysical Research Communications ( IF 2.5 ) Pub Date : 2021-09-20 , DOI: 10.1016/j.bbrc.2021.09.040 Douglas V Laurents 1 , Cristiana Stuani 2 , David Pantoja-Uceda 1 , Emanuele Buratti 2 , Miguel Mompeán 1
The C-terminal, intrinsically disordered, prion-like domain (PrLD) of TDP-43 promotes liquid condensate and solid amyloid formation. These phase changes are crucial to the normal biological functions of the protein but also for its abnormal aggregation, which is implicated in amyotrophic lateral sclerosis (ALS) and certain dementias. We and other previously found that certain amyloid forms emerge from an intermediate condensed state that acts as a nucleus for fibrillization. To quantitatively ascertain the role of individual residues within TDP-43's PrLD in its early self-assembly we have followed the kinetics of NMR 1H–15N HSQC signal loss to obtain values for the lag time, elongation rate and extent of condensate formation at equilibrium. The results of this analysis represent a robust corroboration that aliphatic and aromatic residues are key drivers of condensate formation.
中文翻译:
TDP-43无序区域的芳香族和脂肪族残基正在快速进行自组装
TDP-43 的 C 末端、本质上无序的朊病毒样结构域 (PrLD) 促进液体冷凝物和固体淀粉样蛋白的形成。这些相变对于蛋白质的正常生物学功能至关重要,但对于其异常聚集也是至关重要的,这与肌萎缩侧索硬化症 (ALS) 和某些痴呆症有关。我们和其他人先前发现,某些淀粉样蛋白形式是从中间凝聚状态中出现的,该状态充当纤维化的核。为了定量确定 TDP-43 的 PrLD 中单个残基在其早期自组装中的作用,我们遵循了 NMR 1 H - 15的动力学N HSQC 信号损失以获得滞后时间、伸长率和平衡时冷凝物形成程度的值。该分析的结果有力地证实了脂肪族和芳香族残基是冷凝物形成的关键驱动因素。