Synthesis and Polymerase Recognition of Threose Nucleic Acid Triphosphates Equipped with Diverse Chemical Functionalities
Synthesis and Polymerase Recognition of Threose Nucleic Acid Triphosphates Equipped with Diverse Chemical Functionalities
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DOI：
10.1021/jacs.1c08649
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发表时间：
2021-10-12
影响因子：
15
通讯作者：
Chaput, John C.
中科院分区：
文献类型：
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作者：
Li, Qingfeng;Maola, Victoria A.;Chaput, John C.
扩展可进化非天然遗传聚合物（XNA）的化学空间以包含增强蛋白质靶标结合亲和力的官能团，为具有高生物稳定性的治疗适体提供了一条有前途的途径。在这里，我们描述了在 α-L-苏呋喃糖基尿苷三磷酸 (tUTP) 的 C-5 位引入的 10 个化学不同官能团的化学合成和聚合酶识别。我们证明，tUTP 底物组被实验室进化的聚合酶 Kod-RSGA 普遍识别。从与引物-模板双链体结合的后催化复合物的高分辨率 X 射线晶体结构中获得了对 TNA 合成机制的深入了解。结构分析显示酶活性位点中有一个大空腔，可以容纳 C-5 修饰的 tUTP 底物的侧链。我们的研究结果通过提供人工遗传聚合物的合成路线来扩展可进化核酸系统的化学空间，这些聚合物均经过增强多样性的官能团进行修饰。
Expanding the chemical space of evolvable non-natural genetic polymers (XNAs) to include functional groups that enhance protein target binding affinity offers a promising route to therapeutic aptamers with high biological stability. Here we describe the chemical synthesis and polymerase recognition of 10 chemically diverse functional groups introduced at the C-5 position of alpha-L-threofuranosyl uridine nucleoside triphosphate (tUTP). We show that the set of tUTP substrates is universally recognized by the laboratory- evolved polymerase Kod-RSGA. Insights into the mechanism of TNA synthesis were obtained from a high-resolution X-ray crystal structure of the postcatalytic complex bound to the primer-template duplex. A structural analysis reveals a large cavity in the enzyme active site that can accommodate the side chain of C-5-modified tUTP substrates. Our findings expand the chemical space of evolvable nucleic acid systems by providing a synthetic route to artificial genetic polymers that are uniformly modified with diversity-enhancing functional groups.