Low modularity of aminoacyl-tRNA substrates in polymerization by the ribosome.
Low modularity of aminoacyl-tRNA substrates in polymerization by the ribosome.
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DOI：
10.1093/nar/gkp240
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发表时间：
2009-06
影响因子：
14.9
通讯作者：
Forster AC
中科院分区：
文献类型：
--
作者：
Forster AC
氨基酰基转移rna包含四个标准单元：氨基酸、不变的3 '端CCA、三核苷酸反密码子和tRNA小体。这三个可变模块的互换性程度尚不清楚，尽管它在进化和翻译工程中发挥了作用，以纳入非天然氨基酸。在这里，一个纯化的翻译系统被用来研究各种模块交换对每个肽产物的非天然氨基酰基trna底物的多个核糖体结合效率的影响。含有3到5个相邻的l-氨基酸的非天然侧链产品的产率很低，不能通过优化或简单地用任何单一因素来解释。虽然很容易找到允许定量单一非天然结合的模块组合，但找到能够有效合成含有多种非天然氨基酸的产品的组合是具有挑战性的。这意味着分析多个，而不是单个，每个产品的合并是一个更严格的底物活性测定。大多数结果的不可预测性说明了底物识别的多因素性质和合成生物学对测试我们对翻译的理解的价值。数据表明，氨基酰基trna模块的互换性程度很低。
Aminoacyl-transfer RNAs contain four standardized units: amino acids, an invariant 3′-terminal CCA, trinucleotide anticodons and tRNA bodies. The degree of interchangeability of the three variable modules is poorly understood, despite its role in evolution and the engineering of translation to incorporate unnatural amino acids. Here, a purified translation system is used to investigate effects of various module swaps on the efficiency of multiple ribosomal incorporations of unnatural aminoacyl-tRNA substrates per peptide product. The yields of products containing three to five adjacent l-amino acids with unnatural side chains are low and cannot be improved by optimization or explained simply by any single factor tested. Though combinations of modules that allow quantitative single unnatural incorporations are found readily, finding combinations that enable efficient synthesis of products containing multiple unnatural amino acids is challenging. This implies that assaying multiple, as opposed to single, incorporations per product is a more stringent assay of substrate activity. The unpredictability of most results illustrates the multifactorial nature of substrate recognition and the value of synthetic biology for testing our understanding of translation. Data indicate that the degree of interchangeability of the modules of aminoacyl-tRNAs is low.
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影响因子：
2.9
作者：
Forster, AC;Weissbach, H;Blacklow, SC
通讯作者：
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