期刊
MOLECULAR THERAPY-NUCLEIC ACIDS
卷 13, 期 -, 页码 99-109出版社
CELL PRESS
DOI: 10.1016/j.omtn.2018.08.011
关键词
-
资金
- Department of Biotechnology, Government of India [BT/PR14638/MED/12/483/2010, BT/010/IYBA/2016/10]
- Translational Health Science & Technology Institute, Faridabad
- Indian Council of Medical Research, Government of India
- University Grants Commission, Government of India [22/06/2014 (i) EU-V]
The entry and survival of Mycobacterium tuberculosis (Mtb) within host cells is orchestrated partly by an essential histone- like protein HupB (Rv2986c). Despite being an essential drug target, the lack of structural information has impeded the development of inhibitors targeting the indispensable and multifunctional C-terminal domain (CTD) of HupB. To bypass the requirement for structural information in the classical drug discovery route, we generated a panel of DNA aptamers against HupB protein through systemic evolution of ligands by exponential (SELEX) enrichment. Two G-quadruplex-forming high-affinity aptamers (HupB-4T and HupB-13T) were identified, each of which bound two distinct sites on full-length HupB, with an estimated K-D of similar to 1.72 mu M and similar to 0.17 mu M, respectively, for the high-affinity sites. While HupB-4T robustly inhibited DNA-binding activity of HupB in vitro, both the aptamers recognized surface-located HupB and significantly blocked Mtb entry into THP-1 monocytic cells (p < 0.0001). In summary, DNA aptamers generated in this study block DNA-binding activity of HupB, inhibit virulent Mtb infection in host cells, and demonstrate aptamers to be inhibitors of HupB functions. This study also illustrates the utility of SELEX in developing inhibitors against essential targets for whom structural information is not available.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据