一种设计的 fenylalanyl-tRNA合成酶变体允许有效地在生物体内将aryl ketone功能纳入蛋白质中
Deepshikha Datta1, Pin Wang, Isaac S Carrico
1Division of Chemistry and Chemical Engineering, Howard Hughes Medical Institute, California Institute of Technology, Pasadena, California 91125, USA.
Journal of the American Chemical Society
|May 16, 2002
概括
研究人员通过结合非天然氨基酸来扩大蛋白质合成. 他们使用一种突变酶将p-乙 fenylalanine添加到dihydrofolate reductase中,从而实现化学结合.
科学领域:
- 生物化学和分子生物学
- 合成生物学 合成生物学
- 蛋白质工程是指蛋白质工程.
背景情况:
- 扩大用于蛋白质合成的氨基酸库对于推进蛋白质的设计和功能至关重要.
- 在体内加入非天然氨基酸为创造新型蛋白质功能提供了强大的策略.
- 化学选择性结合使得在温和条件下精确修改蛋白质.
研究的目的:
- 为了证明非天然氨基酸p-乙 fenylalanine在体内被纳入重组蛋白质.
- 为了设计宿主生物体的氨基-tRNA合成酶,以有效地将非天然氨基酸纳入.
- 为了利用纳入的非天然氨基酸用于随后的化学选择性蛋白质结合.
主要方法:
- 来自大肠杆菌的突变基兰-tRNA合成酶的计算设计.
- 在体内将p-乙 fenylalanine纳入重组二叶酸减少酶 (DHFR) 中.
- 基功能化DHFR与化试剂的化学选择性结合.
主要成果:
- 在大肠杆菌中成功将p-乙 fenylalanine纳入重组DHFR.
- 工程合成的fenilalanyl-tRNA合成酶促进了非天然氨基酸的特定充电.
- 在轻度条件下,DHFR上的子功能使得高效的化学选择性结合成为可能.
结论:
- 这项研究建立了一种在体内将基功能化氨基酸纳入蛋白质的方法.
- 开发的方法扩大了蛋白质合成和设计的工具包,使新的功能化策略成为可能.
- 工程蛋白质的化学选择性结合为生物结合和蛋白质修饰提供了一个多功能平台.
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