Department of Chemistry , The Scripps Research Institute , 10550 North Torrey Pines Road , La Jolla , California 92037 , United States.
J Am Chem Soc. 2020 Feb 5;142(5):2125-2128. doi: 10.1021/jacs.9b10538. Epub 2020 Jan 21.
Previously, we evolved a DNA polymerase, SFM4-3, for the recognition of substrates modified at their 2' positions with a fluoro, -methyl, or azido substituent. Here we use SFM4-3 to synthesize 2'-azido-modified DNA; we then use the azido group to attach different, large hydrophobic groups via click chemistry. We show that SFM4-3 recognizes the modified templates under standard conditions, producing natural DNA and thereby allowing amplification. To demonstrate the utility of this remarkable property, we use SFM4-3 to select aptamers with large hydrophobic 2' substituents that bind human neutrophil elastase or the blood coagulation protein factor IXa. The results indicate that SFM4-3 should facilitate the discovery of aptamers that adopt novel and perhaps more protein-like folds with hydrophobic cores that in turn allow them to access novel activities.
此前，我们进化出一种 DNA 聚合酶 SFM4-3，用于识别其 2'位被氟、-甲基或叠氮取代基修饰的底物。在这里，我们使用 SFM4-3 来合成 2'-叠氮修饰的 DNA；然后，我们使用叠氮基团通过点击化学将不同的大疏水性基团连接起来。我们表明，SFM4-3 在标准条件下识别修饰的模板，产生天然 DNA，从而允许扩增。为了证明这一显著特性的实用性，我们使用 SFM4-3 来选择带有大疏水性 2'取代基的适体，这些适体能结合人中性粒细胞弹性蛋白酶或凝血蛋白因子 IXa。结果表明，SFM4-3 应该有助于发现具有新型、可能更类似于蛋白质折叠的适体，这些适体的疏水性核心可以使它们获得新的活性。
