Crystal structures of thrombin in complex with chemically modified thrombin DNA aptamers reveal the origins of enhanced affinity
文献类型：期刊论文
| 作者 | Dolot, Rafal; Lam, Curtis H.; Sierant, Malgorzata; Zhao, Qiang; Liu, Feng-Wu; Nawrot, Barbara; Egli, Martin; Yang, Xianbin |
| 刊名 | NUCLEIC ACIDS RESEARCH
 |
| 出版日期 | 2018-05-18 |
| 卷号 | 46期号:9页码:4819-4830 |
| ISSN号 | 0305-1048 |
| 文献子类 | Article |
| 英文摘要 | Thrombin-binding aptamer (TBA) is a DNA 15-mer of sequence 5 '-GGT TGG TGT GGT TGG-3 ' that folds into a G-quadruplex structure linked by two T-T loops located on one side and a T-G-T loop on the other. These loops are critical for post-SELEX modification to improve TBA target affinity. With this goal in mind we synthesized a T analog, 5-(indolyl-3-acetyl-3-amino-1-propenyl)-2 '-deoxyuridine (W) to substitute one T or a pair of Ts. Subsequently, the affinity for each analog was determined by biolayer interferometry. An aptamer with W at position 4 exhibited about 3-fold increased binding affinity, and replacing both T4 and T12 with W afforded an almost 10-fold enhancement compared to native TBA. To better understand the role of the substituent's aromatic moiety, an aptamer with 5-(methyl-3-acetyl-3-amino1-propenyl)-2 '-deoxyuridine (K; W without the indole moiety) in place of T4 was also synthesized. This K4 aptamer was found to improve affinity 7-fold relative to native TBA. Crystal structures of aptamers with T4 replaced by either W or K bound to thrombin provide insight into the origins of the increased affinities. Our work demonstrates that facile chemical modification of a simple DNA aptamer can be used to significantly improve its binding affinity for a well-established pharmacological target protein. |
| 源URL | [http://ir.rcees.ac.cn/handle/311016/40909]  |
| 专题 | 生态环境研究中心_环境化学与生态毒理学国家重点实验室 |
| 推荐引用方式 GB/T 7714 | Dolot, Rafal,Lam, Curtis H.,Sierant, Malgorzata,et al. Crystal structures of thrombin in complex with chemically modified thrombin DNA aptamers reveal the origins of enhanced affinity[J]. NUCLEIC ACIDS RESEARCH,2018,46(9):4819-4830. |
| APA | Dolot, Rafal.,Lam, Curtis H..,Sierant, Malgorzata.,Zhao, Qiang.,Liu, Feng-Wu.,...&Yang, Xianbin.(2018).Crystal structures of thrombin in complex with chemically modified thrombin DNA aptamers reveal the origins of enhanced affinity.NUCLEIC ACIDS RESEARCH,46(9),4819-4830. |
| MLA | Dolot, Rafal,et al."Crystal structures of thrombin in complex with chemically modified thrombin DNA aptamers reveal the origins of enhanced affinity".NUCLEIC ACIDS RESEARCH 46.9(2018):4819-4830. |
入库方式： OAI收割
来源：生态环境研究中心
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