小分子校正器通过稳定连续折叠状态来使CFTR-F508del从ERAD转移
Celeste Riepe1, Magda Wąchalska1, Kirandeep K Deol2,3,4
1Department of Biology, Stanford University, Stanford, CA, USA 94305.
bioRxiv : the preprint server for biology
|September 25, 2023
概括
囊性纤维化 (CF) 治疗向囊性纤维化跨膜导电性调节器 (CFTR) 蛋白中的F508del突变. 这项研究确定了参与CFTR-F508del降解的关键分子机制,揭示了校正剂如何改善CFTR蛋白水平.
科学领域:
- 分子生物学分子生物学
- 遗传学 遗传学 是一个
- 细胞生物学 细胞生物学
背景情况:
- 超过80%的囊性纤维化 (CF) 患者具有F508del突变,导致错误折叠的囊性纤维化跨膜导电性调节器 (CFTR) 蛋白.
- 错误折叠的CFTR通过内分泌网膜相关降解 (ERAD) 降解,这是当前CF疗法所针对的过程.
- 为了改进校正器的开发,需要对CFTR-F508del ERAD机制有更深入的了解.
研究的目的:
- 通过ERAD系统地确定负责CFTR-F508del降解的分子机制.
- 调查 CFTR-F508del 处理中已识别的因素的作用和小分子校正器的机制.
主要方法:
- 用全基因组的CRISPR/Cas9淘汰屏幕来识别参与CFTR-F508del ERAD.的基因.
- 在RNF5淘汰赛背景下进行了副图书馆选,以识别冗余的连接酶.
- 基因药物相互作用实验使用CFTR校正剂tezacaftor (VX-661) 和elexacaftor (VX-445) 进行.
主要成果:
- 在ER-居民的全方位酶RNF5被确定为一个关键的E3酶,尽管它的淘汰仅部分降低了CFTR-F508del降解.
- 确定RNF185是一种冗余的酶,表明CFTR-F508del ERAD被多个组件缓冲.
- 校正器稳定了CFTR-F508del的RNF5抵抗折叠状态,这表明它们改变了蛋白质的折叠格局.
结论:
- CFTR-F508del ERAD涉及一个缓冲系统,具有冗余的泛素酶,包括RNF5和RNF185.5.
- 小分子校正剂如tezacaftor和elexacaftor似乎通过稳定抗RNF5-介导降解的折叠中间体而起作用.
- 这项工作为CFTR-F508del ERAD提供了洞察力,可能有助于开发更有效的CF疗法.
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