Expanding the library and substrate diversity of the pyrrolysyl-tRNA synthetase to incorporate unnatural amino acids containing conjugated rings.

Expanding the library and substrate diversity of the pyrrolysyl-tRNA synthetase to incorporate unnatural amino acids containing conjugated rings.
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DOI:
10.1002/cbic.201300400
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发表时间:
2013-11-04
期刊:
影响因子:
3.2
通讯作者:
Wang, Lei
Wang, Lei
中科院分区:
生物学3区
文献类型:
--
作者:
Lacey, Vanessa K.;Louie, Gordon V.;Noel, Joseph P.;Wang, Lei

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含有共轭环系统的非天然氨基酸(Uaas)因其光学性质而特别令人感兴趣。到目前为止,这种结构庞大和平面Uaas不能被纳入蛋白质使用吡咯基tRNA/合成酶穿梭系统。通过使用“小智能”方法构建高度多样化的合成酶文库,我们进化出了对两种这样的Uaas特异的新型合成酶,并将它们整合到E.大肠杆菌和哺乳动物细胞。
Unnatural amino acids (Uaas) containing conjugated ring systems are of particular interest for their optical properties. Until now, such structurally bulky and planar Uaas could not be incorporated into proteins using the pyrrolsyl tRNA/synthetase shuttling system. By building a highly diverse synthetase library using the "small intelligent" approach, we evolved novel synthetases specific for two such Uaas and incorporated them into proteins in E. coli and mammalian cells.
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