具有遗传编码"化学弹头"的蛋白质的进化
Chang C Liu1, Antha V Mack, Eric M Brustad
1Department of Chemistry, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, USA.
Journal of the American Chemical Society
|June 27, 2009
概括
我们设计了一种基于菌体的系统,使用非天然的氨基酸进化蛋白质. 纳入p-基氨酸 (BF) 为开发新型糖结合蛋白提供了选择性优势.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 合成生物学 合成生物学
背景情况:
- 基于菌体的系统使蛋白质进化成为可能.
- 扩展的遗传代码允许添加非自然的氨基酸.
- 不自然的氨基酸可以引入新的功能.
研究的目的:
- 研究非自然氨基酸在蛋白质进化中的有用性.
- 为了确定p-boronophenylalanine (BF) 是否在演变中的糖结合蛋白中提供选择性优势.
- 为了证明合成遗传密码在增强蛋白质功能方面的潜力.
主要方法:
- 开发一种基于菌体的系统,用于细菌中的蛋白质进化.
- 加入非天然的氨基酸p-基氨 (BF).
- 使用NNK随机化V(H) CDR3循环选择抗体突变物与模型非循环氨基糖结合.
主要成果:
- 非自然的氨基酸BF在甘氨酸结合蛋白的进化过程中赋予了选择性优势.
- 一个没有偏见的抗体库在被选择与氨基糖结合时聚集在含有BF的突变体上.
- 证明了碳水化合物结合蛋白的成功进化,利用反应性非自然氨基酸.
结论:
- 使用BF的合成遗传密码可以在蛋白质进化中提供选择性优势.
- 这种方法扩大了蛋白质工程可用的功能组谱.
- 代表着向进化碳水化合物结合蛋白具有增强能力的基本步骤.
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