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Full-length TDP-43 and its C-terminal domain form filaments in vitro having non-amyloid properties
Amyloid ( IF 5.5 ) Pub Date : 2020-10-07 , DOI: 10.1080/13506129.2020.1826425
Claudia Capitini 1, 2 , Giulia Fani 1 , Mirella Vivoli Vega 1 , Amanda Penco 3 , Claudio Canale 3 , Lisa D Cabrita 4 , Martino Calamai 2, 5 , John Christodoulou 4 , Annalisa Relini 3 , Fabrizio Chiti 1
Affiliation  

Abstract

Accumulation of ubiquitin-positive, tau- and α-synuclein-negative intracellular inclusions of TDP-43 in the central nervous system represents the major hallmark correlated to amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration with ubiquitin-positive inclusions (FTLD-U). Such inclusions have variably been described as amorphous aggregates or more structured deposits having amyloid properties. Here we have purified full-length TDP-43 (FL TDP-43) and its C-terminal domain (Ct TDP-43) to investigate the morphological, structural and tinctorial features of aggregates formed in vitro by them at pH 7.4 and 37 °C. AFM images indicate that both protein variants show a tendency to form filaments. Moreover, we show that both FL TDP-43 and Ct TDP-43 filaments possess a largely disordered secondary structure, as ascertained by far-UV circular dichroism and Fourier transform infra-red spectroscopy, do not bind Congo red and induce a very weak increase of thioflavin T fluorescence, indicating the absence of a clear amyloid-like signature.



中文翻译:

全长 TDP-43 及其 C 端结构域在体外形成具有非淀粉样蛋白特性的细丝

摘要

中枢神经系统中 TDP-43 泛素阳性、tau 和 α-突触核蛋白阴性细胞内包涵体的积累是与肌萎缩侧索硬化症 (ALS) 和泛素阳性包涵体 (FTLD-U) 的额颞叶变性相关的主要标志。 )。此类内含物被不同地描述为无定形聚集体或更结构化的具有淀粉样蛋白特性的沉积物。在这里,我们纯化了全长 TDP-43 (FL TDP-43) 及其 C 末端结构域 (Ct TDP-43),以研究体外形成的聚集体的形态、结构和色彩特征在 pH 7.4 和 37 °C 下使用它们。AFM 图像表明两种蛋白质变体都显示出形成细丝的趋势。此外,我们表明 FL TDP-43 和 Ct TDP-43 细丝都具有很大程度上无序的二级结构,如远紫外圆二色性和傅里叶变换红外光谱所确定的那样,不结合刚果红并诱导非常微弱的增加硫黄素 T 荧光,表明不存在明显的淀粉样蛋白样特征。

更新日期:2020-10-07
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