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. 2019 Oct 16;9:14867. doi: 10.1038/s41598-019-51244-3

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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PGV-1 inhibits ROS metabolic enzymes. (a) Control, curcumin, and PGV-1 beads were subjected to a pull-down assay using cell lysates containing HA-CBR1, HA-PRDX1, HA-NQO1, HA-NQO2, HA-GLO1, HA-AKR1C1, HA-GSTP1, and HA-GSTO1 proteins. Bound proteins were visualized by immunoblotting using an anti-HA antibody. (b) Docking scores between ROS scavengers and PGV-1 or curcumin. (c) Docking poses between ROS scavengers and PGV-1 (left panels) or curcumin (right panels). Positions of the bound co-factors (FAD, GNB, NADP, and GSH) are also shown. Schematic diagrams of binding between PGV-1 and the corresponding amino acids are shown in middle panels. (d) Competitive pulldown assay between 293T cell lysates containing HA-GSTP1 and curcumin/PGV-1 beads in the presence or absence of 10 mM glutathione. Bound proteins were visualized by immunoblotting using an anti-HA antibody. (e) GST-P1 enzymatic activity was measured in vitro in the presence of curcumin and PGV-1. The IC₅₀ of each compound is shown as the mean ± SD. Km and Vmax were also calculated. (f) K562 cells treated with curcumin (50 μM) and PGV-1 (0.8 μM) for 12, 24 and 48 hr (upper panel), or for 2, 4, and 6 hr (lower panel), were subjected to the ROS detection analysis using FACS.