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. Author manuscript; available in PMC: 2011 May 15.
Published in final edited form as: Biochem J. 2010 Apr 28;428(1):125–131. doi: 10.1042/BJ20100122

Figure 4. The iron-bound hIscA1 acts as an iron donor for the iron-sulfur cluster assembly in E. coli IscU in vitro.

Figure 4

A) Purified E. coli IscU (50 µM) was incubated with E. coli IscS (1 µM), the iron-bound hIscA1 (100 µM), Tris (20 mM, pH 8.0) and NaCl (200 mM) in the presence of dithiothreitol (2 mM) at 37°C for 5 min. L-cysteine (1 mM) was then added to initiate the iron-sulfur cluster assembly reaction. The absorption peak at 456 nm represents formation of the IscU [2Fe-2S] cluster. Spectra were taken every 2 min for 24 min. B) Same as in A) except the iron-bound hIscA1 was replaced with the iron-bound E. coli IscA in the incubation solution.