Department of Nucleic Acids Bioengineering, Institute of Bioorganic Chemistry , Polish Academy of Sciences , Noskowskiego 12/14 , 61-704 Poznan , Poland.
Department of Physics, Chemistry, and Pharmacy, Biomolecular Nanoscale Engineering Center , University of Southern Denmark , Campusvej 55 , Odense M 5230 , Denmark.
J Med Chem. 2019 Mar 14;62(5):2499-2507. doi: 10.1021/acs.jmedchem.8b01806. Epub 2019 Feb 22.
RE31 is a 31-nt DNA aptamer, consisting of the G-quadruplex and a duplex domain, which is able to effectively prolong thrombin time. This article reports on the influence of certain modified nucleotide residues on thermodynamic and biological properties as well as the folding topology of RE31. Particularly, the effect of the presence of nucleosides in unlocked nucleic acid (UNA), locked nucleic acid (LNA), or β-l-RNA series was evaluated. The studies presented herein show that all modified residues can influence thermal and biological stabilities of G-quadruplex in a position-dependent manner. The aptamers modified simultaneously with UNA at the T position and LNAs in the duplex part possess the highest value of melting temperature and a 2-fold higher anticoagulant effect. Importantly, RE31 variants modified with nucleosides in UNA, LNA, or β-l-RNA series exhibit unchanged G-quadruplex folding topology. Crucially, introduction of any of the modified residues into RE31 causes prolongation of aptamer stability in human serum.
RE31 是一个由 31 个核苷酸组成的 DNA 适体，由 G-四链体和双链区组成，能够有效地延长凝血酶时间。本文报道了某些修饰核苷酸残基对 RE31 的热力学和生物学性质以及折叠拓扑结构的影响。特别是，评估了非锁核酸（UNA）、锁核酸（LNA）或β-L-RNA 系列中核苷的存在对 RE31 的影响。本文所述的研究表明，所有修饰的残基都可以以位置依赖的方式影响 G-四链体的热稳定性和生物学稳定性。在 T 位置同时用 UNA 修饰和在双链部分用 LNA 修饰的适体具有最高的熔点值和 2 倍更高的抗凝效果。重要的是，用 UNA、LNA 或β-L-RNA 系列中的核苷修饰的 RE31 变体保持不变的 G-四链体折叠拓扑。至关重要的是，将任何修饰的残基引入 RE31 都会导致适体在人血清中的稳定性延长。
