在体内鉴定二硫化键形成所需的蛋白质
J C Bardwell1, K McGovern, J Beckwith
1Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115.
Cell
|November 1, 1991
概括
在dsbA基因的突变损害了Escherichia coli中二硫化键的形成. DsbA蛋白在体内促进二硫化物键的产生,这对于适当的蛋白质折叠至关重要.
科学领域:
- 微生物学 微生物学
- 分子生物学分子生物学
- 蛋白质生物化学 蛋白质生物化学
背景情况:
- 二硫化物键对于许多分泌的蛋白质的稳定性和功能至关重要.
- 大肠杆菌依赖于在周围质中有效形成二硫化物键的特定机制.
研究的目的:
- 研究dsbA基因在大肠杆菌中二硫化键形成中的作用.
- 描述DsbA蛋白及其在蛋白质折叠中的功能.
主要方法:
- 在大肠杆菌中dsbA突变的遗传分析.
- 脉冲追踪标记和蛋白酶敏感性测定用于分泌的蛋白质.
- 净化DsbA蛋白活性的生物化学特征.
主要成果:
- dsbA突变导致分泌蛋白质的二硫化键形成严重缺陷,如β-乳酸酶,性酸酶和OmpA.
- 在dsbA突变体中缺乏二硫化键的蛋白质似乎是折叠中间体,表现出蛋白酶敏感性.
- dsbA基因编码了一个周等离子蛋白 (DsbA),其活性位点与二硫化氧化还原酶相同.
- 纯化DsbA蛋白显示出降低胰岛素中二硫化键的能力.
结论:
- DsbA蛋白对于促进大肠杆菌中二硫化键的形成至关重要.
- DsbA可能在细菌周围质中作为氧化蛋白折叠通路中的关键酶起作用.
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