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NIFS and H. influenzae selenophosphate synthetase coupled assays. Assays were performed anaerobically at 37°C in 100 mM Tricine⋅KOH (pH 8.0), 2 mM DTT, 20 mM KCl, 4 mM MgCl2, 2 mM ATP, 0.2 μCi [14C]ATP, 2 mM l-selenocysteine, 5 μM selenophosphate synthetase, and the indicated concentration of NIFS.
