Pyrrolysyl-tRNA synthetase: an ordinary enzyme but an outstanding genetic code expansion tool.

Pyrrolysyl-tRNA synthetase: an ordinary enzyme but an outstanding genetic code expansion tool.
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DOI:
10.1016/j.bbapap.2014.03.002
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发表时间:
2014-06
影响因子:
3.2
通讯作者:
Liu, Wenshe R.
Liu, Wenshe R.
中科院分区:
生物学3区
文献类型:
--
作者:
Wan, Wei;Tharp, Jeffery M.;Liu, Wenshe R.

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琥珀密码子处第22个蛋白质氨基酸pyrolysine(Pyl)的遗传掺入是通过吡咯赖氨酰-tRNA合成酶(PylRS)及其同源tRNAPyl的作用实现的。与大多数氨酰-tRNA合成酶不同,PylRS显示出高底物侧链混杂性、对其底物α-胺的低选择性和对tRNAPyl的反密码子的低选择性。PylRS作为氨酰-tRNA合成酶的这些独特但普通的特征允许Pyl掺入机器容易地被工程化以用于在琥珀密码子处将超过100个非经典氨基酸(NCAA)或α-羟基酸遗传掺入到蛋白质中,以及重新分配其他密码子如赭石UAA、蛋白石UGA和四碱基AGGA密码子以编码NCAA。
The genetic incorporation of the 22nd proteinogenic amino acid, pyrolysine (Pyl) at amber codon is achieved by the action of pyrrolysyl-tRNA synthetase (PylRS) together with its cognate tRNAPyl. Unlike most aminoacyl-tRNA synthetases, PylRS displays high substrate side chain promiscuity, low selectivity toward its substrate α-amine, and low selectivity toward the anticodon of tRNAPyl. These unique but ordinary features of PylRS as an aminoacyl-tRNA synthetase allow the Pyl incorporation machinery to be easily engineered for the genetic incorporation of more than 100 non-canonical amino acids (NCAAs) or α-hydroxy acids into proteins at amber codon and the reassignment of other codons such as ochre UAA, opal UGA, and four-base AGGA codons to code NCAAs.
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