含有非折形螺旋域的复合指的设计和结构确定
Caterina Maria Lombardo1, Vasantha Kumar M V2, Céline Douat1
1Univ. Bordeaux, CNRS, CBMN, UMR 5248, Institut Européen de Chimie et Biologie , 2 rue Robert Escarpit , 33607 Pessac , France.
Journal of the American Chemical Society
|February 5, 2019
概括
研究人员通过用合成折叠剂替换指中的天然α螺旋来制造人造蛋白质. 这种工程化蛋白质混合物保持协调和DNA结合,为新生物材料铺平了道路.
科学领域:
- 生物化学
- 合成生物学
- 蛋白质工程
背景情况:
- 折叠体模仿蛋白质的二次结构,使得特定蛋白质相互作用的合成寡合体设计.
- 合成的骨干可以替代蛋白质中的天然片,有可能创造具有新功能的人工蛋白质.
- 橄素2. 5螺旋体与类α螺旋体具有结构相似性,这表明在保持三级结构的同时可以替换脊柱.
研究的目的:
- 通过将Cys2His2指中的α-螺旋段替换为酸氨酸折叠剂来制造复合蛋白.
- 评估工程指混合物是否保留结合能力和原生结构.
- 评估修改后的指的DNA结合活性.
主要方法:
- 将Zif268指3中的原生α螺旋替换为含有伊米达的氨酸序列.
- 使用光谱技术和质谱测量进行原生条件分析.
- 使用核磁共振 (NMR) 光谱进行详细的结构分析.
- 通过对Egr1目标DNA序列进行定位DNA结合测试.
主要成果:
- /氨酸混合物成功地协调了离子,不受折叠剂替代的影响.
- 核磁共振分析证实工程指采用了折叠结构.
- 保存了原生β片排列和DNA结合侧链方向.
- 经过修改的指与野生类型类似地与GC基结合.
结论:
- 替换一个指的阿尔法螺旋段与一个酸折叠器是可行的.
- 工程复合蛋白保留了重要的功能,包括协调和DNA结合.
- 这项研究代表了用设计的折叠元件构建人工蛋白质的重要一步.
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