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. 2018 Jul 24;9:1659. doi: 10.3389/fmicb.2018.01659

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Copyright © 2018 García, Carreño, Tirado-Vélez, Ferrándiz, Rodrigues, Gracia, Amblar, Ainsa and de la Campa.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Inhibition of M. tuberculosis topoisomerase I by SCN and N-SCN. (A,B) Supercoiled pBR322 (0.5 μg) was treated with 1 unit of purified MtbTopoI in 200 μl reactions containing N-SCN, SCN, or boldine at the concentrations (μM) indicated. Mw, molecular mass standard; S, covalently closed supercoiled pBR322 used as substrate. (C) One unit of MtbTopoI was pre-incubated with SCN at the indicated concentrations, then 0.1 pmol of 5′-biotin-labeled 32-mer oligonucleotide was added and the reaction was continued for 30 min. The products were resolved in 12% denaturing PAGE. (D) Quantification of the 19-mer cleavage product. The values (mean ± SD) of three independent experiments are shown. ^∗ indicates statistical significance by Student T-test P < 0.05.