RIBOSOME-MEDIATED INCORPORATION OF A NONSTANDARD AMINO-ACID INTO A PEPTIDE THROUGH EXPANSION OF THE GENETIC-CODE
RIBOSOME-MEDIATED INCORPORATION OF A NONSTANDARD AMINO-ACID INTO A PEPTIDE THROUGH EXPANSION OF THE GENETIC-CODE
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DOI：
10.1038/356537a0
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发表时间：
1992-04-09
期刊：
影响因子：
64.8
通讯作者：
BENNER, SA
中科院分区：
文献类型：
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作者：
BAIN, JD;SWITZER, C;BENNER, SA
蛋白质工程师面临的一个严重局限性是只有20'蛋白质原性的氨基酸，由天然信使RNA编码。这些氨基酸之间缺乏结构多样性限制了可以通过位置定向的诱变来解决的机械和结构问题。在这里，我们描述了一种通过基于核糖体的翻译将非标准氨基酸掺入多肽中的新技术。在这项技术中，通过从具有非标准氢键模式的非天然核苷基碱基中创建第65个密码子 - 抗病年对通过创建第65个密码子 - 抗毒剂对的扩展。这种新的密码子 - 抗原对有效地支持体外翻译，以产生含有非标准氨基酸的肽。核糖体作为合成工具的多功能性为蛋白质工程提供了新的可能性，并与另一种最近描述的方法进行了比较，在该方法中，遗传密码被简单地重新排列以募集终止密码子以3-9扮演编码角色。
ONE serious limitation facing protein engineers is the availability of only 20 'proteinogenic' amino acids encoded by natural messenger RNA. The lack of structural diversity among these amino acids restricts the mechanistic and structural issues that can be addressed by site-directed mutagenesis. Here we describe a new technology for incorporating non-standard amino acids into polypeptides by ribosome-based translation. In this technology, the genetic code is expanded through the creation of a 65th codon-anticodon pair from unnatural nucleoside bases having non-standard hydrogen-bonding patterns 1,2. This new codon-anticodon pair efficiently supports translation in vitro to yield peptides containing a non-standard amino acid. The versatility of the ribosome as a synthetic tool offers new possibilities for protein engineering, and compares favourably with another recently described approach in which the genetic code is simply rearranged to recruit stop codons to play a coding role 3-9.