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. Author manuscript; available in PMC: 2018 Oct 31.
Published in final edited form as: Biochemistry. 2017 Oct 19;56(43):5726–5738. doi: 10.1021/acs.biochem.7b00781

Figure 3.

Figure 3

Analytical HPLC traces (220 nm absorption) from enzymatic activity assays employing the mammalian thioredoxin system and S100A7ox as a substrate. Conditions: 5 μM S100A7ox, 1 μM human Trx, 0.1 μM rat liver TrxR, and 1 mM NADPH (75 mM HEPES, 100 mM NaCl, pH 7.0, 37 °C) with (A) no metal added, (B) no enzyme added, (C) 2 mM Ca(II) added to the buffer, (D) 1.9 equiv of Zn(II) added, and (E) 1.9 equiv of Zn(II) and 2 mM Ca(II) added to the buffer. In each panel, the bottom chromatogram is a S100A7red standard. Data from control assays are presented in Figures S6, S8, and S9. The shoulder peaks observed in the chromatograms of S100A7ox and S100A7red correspond to S100A7 isoforms missing the N-terminal methionine.