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. Author manuscript; available in PMC: 2015 Aug 1.
Published in final edited form as: Mol Microbiol. 2014 Jul 8;93(4):629–644. doi: 10.1111/mmi.12676

Figure 4. The conserved cysteine residues are essential for the copper binding in IscA.

Figure 4

A), UV-visible spectra of IscA and the IscA mutant (IscA-3M) after incubation with iron in vitro. Apo-IscA and the IscA triple mutant (50 μM dimer) were incubated with Fe(NH4)2 (SO4)2 (100 μM) in the presence of dithiothreitol (2 mM) at room temperature for 15 min, followed by protein re-purification. B), UV-visible spectra of IscA and the IscA mutant (IscA-3M) after incubation with copper in vitro. Apo-IscA and the IscA mutant (50 μM dimer) were incubated with CuSO4 (100 μM) in the presence of dithiothreitol (2 mM) at room temperature for 15 min, followed by protein re-purification. C), UV-visible spectra of IscA and the IscA mutant (IscA-3M) purified from the E. coli copA/cueO/cusA mutant cells grown in LB media supplemented with Fe(NH4)2 (SO4)2 (200 μM). D), UV-visible spectra of IscA and the IscA mutant (IscA-3M) purified from the E. coli copA/cueO/cusA mutant cells grown in LB media supplemented with CuSO4 (200 μM).