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8-17 DNA 酶的可编程催化核心。

The Programmable Catalytic Core of 8-17 DNAzymes.

机构信息

School of Pharmaceutical Sciences, Guizhou University, Guiyang 550025, China.

Beijing Institute of Pharmacology and Toxicology, Taiping 27, Beijing 100850, China.

出版信息

Molecules. 2024 May 21;29(11):2420. doi: 10.3390/molecules29112420.

Abstract

8-17 DNAzymes (8-17, 17E, Mg5, and 17EV1) are in vitro-selected catalytic DNA molecules that are capable of cleaving complementary RNAs. The conserved residues in their similar catalytic cores, together with the metal ions, were suggested to contribute to the catalytic reaction. Based on the contribution of the less conserved residues in the bulge loop residues (W, A, A) and the internal stem, new catalytic cores of 8-17 DNAzymes were programmed. The internal stem CTC-GAG seems to be more favorable for the DNAzymes than CCG-GGC, while an extra W led to a significant loss of activity of DNAzymes, which is contrary to the positive effect of A, by which a new active DNAzyme 17EM was derived. It conducts a faster reaction than 17E. It is most active in the presence of Pb, with the metal ion preference of Pb >> Zn > Mn > Ca ≈ Mg. In the Pb and Zn-mediated reactions of 17EM and 17E, the same Na- and pH dependence were also observed as what was observed for 17E and other 8-17 DNAzymes. Therefore, 17EM is another member of the 8-17 DNAzymes, and it could be applied as a potential biosensor for RNA and metal ions.

摘要

8-17DNA 酶(8-17、17E、Mg5 和 17EV1)是体外筛选出的具有切割互补 RNA 能力的催化 DNA 分子。其相似催化核心中的保守残基与金属离子共同促进催化反应。基于茎环区(W、A、A)和内部茎中不那么保守的残基的贡献,设计了新的 8-17DNA 酶催化核心。内部茎 CTC-GAG 似乎比 CCG-GGC 更有利于 DNA 酶,而额外的 W 则导致 DNA 酶活性显著丧失,这与 A 的积极作用相反,由此产生了新的活性 DNA 酶 17EM。它的反应速度比 17E 更快。在 Pb 的存在下,它具有最高的活性,金属离子的偏好为 Pb >> Zn > Mn > Ca ≈ Mg。在 17EM 和 17E 的 Pb 和 Zn 介导反应中,也观察到了与 17E 和其他 8-17DNA 酶相同的 Na+和 pH 依赖性。因此,17EM 是 8-17DNA 酶的另一个成员,可作为 RNA 和金属离子的潜在生物传感器。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0d8e/11173380/84a9a554794f/molecules-29-02420-g001.jpg

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