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. 2016 Feb 25;39(3):242–249. doi: 10.14348/molcells.2016.2271

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Fig. 3. — Enzyme activity and binding affinity of an inhibitor (DDC) of hEC-SOD^f and hEC-SOD^tr in the presence of 0.1% BSA & 50 μM Cu/Zn. (A) Activity assay of hEC-SOD^f in the presence of 0.1% BSA, and 50 μM Cu/Zn ions. (B) Activity assay of hEC-SOD^tr in the presence of 0.1% BSA, and 50 μM Cu/Zn ions. (C) Fluorescence assay of hEC-SOD^f with inhibitor (Na-DDC) in the presence of 0.1% BSA; 50 μM Cu/Zn. (D) Fluorescence assay of hEC-SOD^tr with inhibitor (Na-DDC) in the presence of 0.1% BSA; 50 μM Zn/Cu. Protein samples were excited at 280 nm and emission spectra were recorded from 300 to 450 nm. All the assays were performed in phosphate-buffered saline (PBS; pH 7.4) using 20 μM hEC-SOD with an increasing concentration of inhibitor (Na-DDC). [ 0 μM; 10 μM; 20 μM; 30 μM; 40 μM; 50 μM; 60 μM; 70 μM; 80 μM; 90 μM and 100 μM]. (E) Effect of Na-DDC (inhibitor) on hEC-SOD activity. Enzyme activity of hEC-SOD was determined using the SOD Assay Kit WST-1 (Dojindo Laboratories) in PBS buffer (pH 7.4) in the absence and presence of 50 μM of inhibitor (Na-DDC).

Inline graphic — Enzyme activity and binding affinity of an inhibitor (DDC) of hEC-SOD^f and hEC-SOD^tr in the presence of 0.1% BSA & 50 μM Cu/Zn. (A) Activity assay of hEC-SOD^f in the presence of 0.1% BSA, and 50 μM Cu/Zn ions. (B) Activity assay of hEC-SOD^tr in the presence of 0.1% BSA, and 50 μM Cu/Zn ions. (C) Fluorescence assay of hEC-SOD^f with inhibitor (Na-DDC) in the presence of 0.1% BSA; 50 μM Cu/Zn. (D) Fluorescence assay of hEC-SOD^tr with inhibitor (Na-DDC) in the presence of 0.1% BSA; 50 μM Zn/Cu. Protein samples were excited at 280 nm and emission spectra were recorded from 300 to 450 nm. All the assays were performed in phosphate-buffered saline (PBS; pH 7.4) using 20 μM hEC-SOD with an increasing concentration of inhibitor (Na-DDC). [ 0 μM; 10 μM; 20 μM; 30 μM; 40 μM; 50 μM; 60 μM; 70 μM; 80 μM; 90 μM and 100 μM]. (E) Effect of Na-DDC (inhibitor) on hEC-SOD activity. Enzyme activity of hEC-SOD was determined using the SOD Assay Kit WST-1 (Dojindo Laboratories) in PBS buffer (pH 7.4) in the absence and presence of 50 μM of inhibitor (Na-DDC).