重组性Trichomonas vaginalis 20S蛋白酶与共价抑制剂结合的结构阐明
Jan Silhan1, Pavla Fajtova1,2, Jitka Bartosova1
1Institute of Organic Chemistry and Biochemistry AS CR, v.v.i., Flemingovo nam. 2., 166 10 Prague 6, Czech Republic.
bioRxiv : the preprint server for biology
|August 30, 2023
概括
研究人员为结构研究开发了一种复合的Trichomonas vaginalis 20S蛋白酶体 (Tv20S). 这种Tv20S被marizomib和carmaphycin-17抑制,为新的三病治疗提供了基础.
科学领域:
- 生物化学 生物化学
- 结构生物学 结构生物学
- 寄生虫学的寄生虫学
背景情况:
- 20S蛋白酶对真核生物中的蛋白质稳定至关重要,包括原生虫寄生虫Trichomonas vaginalis (Tv).
- 电视蛋白质体是治疗三虫病的有效药物标,但本地酶研究受到低产量和纯度的阻碍.
- 再组合表达是克服这些限制的可行策略,用于生化和结构研究.
研究的目的:
- 为了产生适合生物化学和结构分析的重组Tv 20S蛋白质组.
- 通过已知的蛋白酶体抑制剂,marizomib (MZB) 和carmaphycin-17 (CP-17) 来研究Tv20S的抑制.
- 使用冷电子显微镜 (cryo-EM) 阐明抑制剂与Tv20S结合的结构基础.
主要方法:
- 在昆虫细胞中表达Tv 20S蛋白质组的所有七个α和七个β子单元以及Ump-1伴侣.
- 重组Tv20S蛋白酶的净化和生物化学表征.
- 同结晶和冷电子显微镜 (cryo-EM) 用于确定Tv20S与MZB和CP-17复合的结构.
主要成果:
- 成功产生了一种重组的Tv20S蛋白酶体,该蛋白质体在生物化学上与原生酶相同.
- 证实MZB和CP-17抑制重组Tv20S,具有不同的结合位点.
- 获得了高分辨率 (2.8Å) 的冷电磁结构,揭示了Tv20S折叠和抑制剂结合模式.
- 解释了MZB和CP-17与TV20S相比的人类蛋白质组的差异特异性.
结论:
- 重组Tv20S蛋白质组作为结构和生化研究的强大平台.
- 对抑制剂结合的结构洞察力为开发向的Tv20S抑制剂提供了基础.
- 这项研究为通过准TV蛋白质组而对抗三虫病的新型治疗策略铺平了道路.
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