Issue 44, 2014

Stability and bioactivity of thrombin binding aptamers modified with d-/l-isothymidine in the loop regions

Abstract

Thrombin binding aptamer (TBA) is a 15-mer single-strand DNA that was identified by SELEX screening technology. It adopts a chair-type antiparallel G-quadruplex and can specifically interact with thrombin, thus inhibiting blood coagulation. Isonucleoside (isoNA) is a type of nucleoside isomer in which the base is shifted to 2′-positions of the glycosyl group, endowed with the ability to modulate local conformation of nucleotides, and L-isoNA could alter the conformation more due to the inversion of glycosyl configuration. Incorporation of L-isothymidine (L-isoT) at T3, T9, T12 positions and D-isoT at the T7 position in TBA's loop regions promoted the formation of G-quadruplex, resulting in enhanced affinity with thrombin and an increased anticoagulant effect. Computer simulation indicated that TBA-12L showed the strongest binding with thrombin, which was consistent with experimental results. The bioactivity of double isoNA incorporated TBA with D-IsoT at T7 and L-IsoT at T12 was comparable to that of TBA-12L, suggesting that the T12 of TBA was very important in interaction with thrombin. Our study also suggested that TBA might interact with two thrombin molecules through the TT loops (T3T4, T12T13) and TGT loop, but the second bonding did not show additional biological effects.

Graphical abstract: Stability and bioactivity of thrombin binding aptamers modified with d-/l-isothymidine in the loop regions

Supplementary files

Article information

Article type
Paper
Submitted
19 Jul 2014
Accepted
09 Sep 2014
First published
09 Sep 2014

Org. Biomol. Chem., 2014,12, 8866-8876

Author version available

Stability and bioactivity of thrombin binding aptamers modified with D-/L-isothymidine in the loop regions

B. Cai, X. Yang, L. Sun, X. Fan, L. Li, H. Jin, Y. Wu, Z. Guan, L. Zhang, L. Zhang and Z. Yang, Org. Biomol. Chem., 2014, 12, 8866 DOI: 10.1039/C4OB01525H

To request permission to reproduce material from this article, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements