一个基因编码的aza-Michael受体,用于蛋白质受体复合体的共价交叉链接
Jennifer L Furman1, Mingchao Kang, Seihyun Choi
1Department of Chemistry and the Skaggs Institute for Chemical Biology, The Scripps Research Institute , 10550 North Torrey Pines Road, La Jolla, California 92037, United States.
Journal of the American Chemical Society
|May 22, 2014
概括
研究人员设计了非自然的氨基酸,在蛋白质接口上产生选择性的共价键. 这种方法可以实现精确的蛋白质交叉链接,为新的生物探针和疗法提供了潜力.
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:
- 向对蛋白质的共价修饰具有挑战性.
- 基因编码非自然氨基酸为特定位点的蛋白质功能提供了一条途径.
研究的目的:
- 进化直角遗传系统,以结合具有电友性质的非自然氨基酸.
- 用这些工程氨基酸在蛋白质-蛋白质接口上演示选择性共价键的形成.
主要方法:
- 正交氨基酸-tRNA合成酶/tRNACUA对的演变.
- 在大肠杆菌中对三个aza-Michael受体氨基酸 (AcrK,AcrF,VSF) 的遗传编码.
- 使用αErbB2 Fab-ErbB2抗体-受体系统来证明交叉链接.
主要成果:
- 在大肠杆菌中成功编码非自然氨基酸.
- 在alphaErbB2-VSF突变体和ErbB2的氨酸残留物之间展示了特定位置的共价交叉链接.
- 在体外在生理pH下,在体外实现了净化和细胞ErbB2的近量交叉链接.
结论:
- 开发了一种遗传编码非自然氨基酸的方法,这些氨基酸形成有选择性的共价键.
- 这种方法可以实现高效和生物相容的蛋白质交叉链接.
- 潜在的应用包括新的细胞生物探针,诊断和治疗.
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