Figure 4.
Gel-filtration analysis of AF and HMAF-inactivated TrxR. TrxR (80 nM) was first incubated with NADPH (100 μM) at 25 °C for 10 min followed by the addition of test compounds and further incubation for 2 h at 25 °C (A, AF, 3.3, 6.7, 16.7, 33.3, 66.7 μM; B, HMAF, 0.13, 0.27, 0.53, 1.33, 2.67 μM). Unbound compound was removed by Micro Bio-Spin™ P-6 pre-packed size exclusion columns. The residual activity was measured with DTNB assay described in the experimental details.