一个内在失调的-接器,用于高效的蛋白质结合,无论是在体外还是体外
Xia-Ling Wu1, Yajie Liu1, Dong Liu1
1Key Laboratory of Polymer Chemistry and Physics of Ministry of Education, Center for Soft Matter Science and Engineering, College of Chemistry and Molecular Engineering , Peking University , Beijing 100871 , P. R. China.
Journal of the American Chemical Society
|November 20, 2018
概括
我们开发了SpyStapler, 这是一种无序的蛋白质, 能有效催化SpyTag和BDTag之间的同位素键形成. 这使得体外和体内蛋白质结合能够产生复杂的蛋白质结构.
科学领域:
- 蛋白质工程
- 生物化学
- 分子生物学
背景情况:
- 通过SpyCatcher和SpyTag形成一个高效的蛋白质结合系统.
- 现有的蛋白质结合方法可能受到反应条件和效率的限制.
研究的目的:
- 设计一种新型的蛋白质连接酶SpyStapler,
- 证明SpyStapler在体外和体内蛋白质生物结合和循环蛋白质合成中的有用性.
主要方法:
- 使用SpyCatcher的蛋白质工程来创建SpyStapler和BDTag.
- 在体外和体内蛋白质结合试验.
- 循环二重化和核磁共振光谱用于结构和机械研究.
主要成果:
- 在SpyTag和BDTag之间SpyStapler有效催化异键的形成,在体外产量为80%左右.
- 通过联合表达实现了循环二酸减少酶 (DHFR) 的细胞合成.
- 在标签的结合和绑定时,SpyStapler从无序转变为稳定的折叠状态,异键稳定复合体.
结论:
- 它是蛋白质生物结合的强大而有效的工具.
- 对于SpyStapler的催化作用来说,它的无序性和过渡到有序状态至关重要.
- 提供了构建复杂蛋白质架构和合成生物学应用的新可能性.
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