Flexizyme as a versatile tRNA acylation catalyst and the application for translation.
Flexizyme as a versatile tRNA acylation catalyst and the application for translation.
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DOI：
10.1093/nass/nrl018
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发表时间：
2006-01-01
期刊：
影响因子：
--
通讯作者：
Suga, Hiroaki
中科院分区：
文献类型：
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作者：
Murakami, Hiroshi;Ohta, Atushi;Suga, Hiroaki
在这里，我们描述了一个从头tRNA酰化系统组成的人工核酶。该系统，flexizyme（Fx）系统，允许制备酰基-tRNA，对各种氨基酸/羟基酸和tRNA的使用几乎没有限制。为了证明它的效用，我们已经进行了蛋白质合成，包括通过琥珀或程序移码辅助位点特异性掺入非天然氨基酸和羟基酸。我们还证明了通过有义密码子抑制涉及多个非天然氨基酸的肽合成。Fx系统和适当的无细胞翻译的组合是用于mRNA编码的非天然多肽合成的强大且灵活的工具。
Here we describe a de novo tRNA acylation system consisting of artificial ribozymes. This system, the flexizyme (Fx) system, allows for the preparation of acyl-tRNAs with almost no limitation on the use of a variety of amino/hydroxy acids and tRNAs. To demonstrate its utility, we have carried out protein synthesis involving site-specific incorporation of nonnatural amino and hydroxy acids via amber or programmed frame-shift suppressions. We have also demonstrated peptide synthesis involving multiple nonnatural amino acids via sense codon suppression. The combination of the Fx system and appropriate cell-free translation is a powerful and flexible tool for mRNA-encoded synthesis of nonnatural polypeptides.