In vitro selection of ATP-binding receptors using a ribonucleopeptide complex.
In vitro selection of ATP-binding receptors using a ribonucleopeptide complex.
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使用核糖核酸肽复合物体外选择 ATP 结合受体。
DOI：
10.1021/ja016569x
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发表时间：
2002
影响因子：
15
通讯作者：
K. Makino
中科院分区：
文献类型：
--
作者：
T. Morii;M. Hagihara;Shin;K. Makino
最近描述的核糖体的三维结构提供了一种RNA-蛋白质复合体的显着多样性的感觉。我们设计了一类新型的人工受体支架，在支架中，一个短肽与具有随机核苷酸区域的RNA形成稳定而特异的复合体。随机的核苷酸区域被放置在HIV-1REV反应元件旁边，以便在REV肽存在的情况下形成“核糖核肽池”。在体外，从随机库中选择RNA寡核苷酸提供了一种对ATP具有特异性的核糖核肽受体。与ATP结合的核糖核肽与已知的ATP适配子的核苷酸序列不一致，在没有REV肽的情况下完全失去了与ATP结合的能力。通过取代REV多肽的N端氨基酸，提高了核糖核肽的ATP结合活性。这些结果直接证明了该肽被掺入到RNA的功能结构中，并表明该肽的RNA结合区以外的氨基酸调节了核糖核肽的ATP结合。我们的方法将为“定制”核糖核肽受体和酶的设计提供另一种策略。
A recently described three-dimensional structure of the ribosome provides a sense of remarkable diversity of RNA-protein complexes. We have designed a new class of scaffold for artificial receptors, in which a short peptide and RNA with a randomized nucleotide region form a stable and specific complex. The randomized nucleotide region was placed next to the HIV-1 Rev response element to enable the formation of "ribonucleopeptide" pools in the presence of the Rev peptide. In vitro selection of RNA oligonucleotides from the randomized pool afforded a ribonucleopeptide receptor specific for ATP. The ATP-binding ribonucleopeptide did not share the known consensus nucleotide sequence for ATP aptamers and completely lost its ATP-binding ability in the absence of the Rev peptide. The ATP-binding activity of the ribonucleopeptide was increased by a substitution of the N-terminal amino acid of the Rev peptide. These results demonstrate directly that the peptide is incorporated in the functional structure of RNA and suggest that amino acids outside the RNA-binding region of the peptide modulate the ATP-binding of ribonucleopeptide. Our approach would provide an alternative strategy for the design of "tailor-made" ribonucleopeptide receptors and enzymes.