染色体9的自动化快速流合成 72个二重复蛋白
Kohei Sato1,2, Charlotte E Farquhar2, Jacob Rodriguez2
1School of Life Science and Technology, Tokyo Institute of Technology, 4259 Nagatsuta-cho, Midori-ku, Yokohama-shi, Kanagawa 226-8501, Japan.
Journal of the American Chemical Society
|June 9, 2023
概括
在C9orf72中,六核酸重复扩张会通过产生有毒的二酸重复蛋白 (DPR) 来引起ALS和FTD. 这项研究合成了DPR,揭示了结构性质和细胞毒性,有助于疾病模型的发展.
科学领域:
- 神经科学
- 遗传学
- 生物化学
背景情况:
- C9orf72重复扩张是ALS和FTD的主要遗传原因.
- 这种突变产生涉及神经退行症的有毒二重复蛋白 (DPR).
- DPRs的有限可用性阻碍了对其特性和病原机制的研究.
研究的目的:
- 用于化学合成c9orf72二重复蛋白 (DPR).
- 描述合成DPR的物理化学特性.
- 评估神经元细胞中合成DPR的细胞毒性.
主要方法:
- 自动化快流合成 (AFPS) 用于DPR合成 (多达200个氨基酸).
- 循环二色谱法以确定二次结构.
- 用于聚合分析的尺寸排除色谱.
- 使用人类神经母细胞的细胞活力测试.
主要成果:
- 含有proline的DPR (多PR,多GP,多PA) 采用了类似于polyproline II的螺旋结构.
- 较长的多GP和多PA显示了聚合的潜力.
- 复制时间较长的多GR和多PR降低了细胞活力,模仿了内源性DPR细胞毒性.
- 多GP和多PA没有显著的细胞毒性.
结论:
- AFPS有效合成像DPRs这样的低复杂性.
- 合成DPR具有与内源性DPR相似的结构和细胞毒性.
- 这项工作有助于研究DPR的致病机制,并开发ALS和FTD的疾病模型.
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