Fig. 4. The putative zinc binding site at the dimer interface.

a Structural details of the zinc binding site. b Sequence alignment of the CXXC-containing insertion between NOX5 paralogs and orthologs. The CXXC motif is indicated with black line and black box in left and right panel, respectively. c ICP-MS shows the presence of Zinc ions in NOX5. In x-axis: buffer indicates NOX5 sample buffer (20 mM Tris, 200 mM NaCl, 0.05% Digitonin); filtrate indicates the buffer passed through the filter device during NOX5 concentration; NOX5 is the experimental group. Data are shown with n = 3 technical repeats. Source data are provided as a Source Data file. d Relative activity of C571S and C568S to wild-type NOX5. Y-axis indicates the relative activity compared with NOX5 wild type. Data are shown (average +/-SD) with n = 5 biologically independent samples. Source data are provided as a Source Data file. e Dose-dependent inhibition of NOX5 by TPEN. Y-axis indicates the relative activity compared with NOX5 wild type. Data are shown with (average +/-SD) n = 5 biologically independent samples. Data are processed with GraphPad Prism (Version 10.1.1). We performed Grubbs’ test⁵⁹ (also known as extreme studentized deviate, ESD), to determine the outliers. We excluded one data point in “2 mM TPEN” group, which is a significant outliner (P < 0.05) based on Grubbs’ test. Source data are provided as a Source Data file.