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Ag(I), Hg(II), Cd(II), and Zn(II) also damaged the [4Fe-4S] clusters of fumarase A in vivo. Strain SJ32 (0.2 OD unit) was exposed to soft metals for 30 min in glucose medium in the presence of chloramphenicol to block protein synthesis. Lysates were then prepared and analyzed in anaerobic buffer. Lysates were assayed without further treatment, after incubation with Fe(II)-DTT, or after incubation with Fe(II)-DTT-IscS-cysteine.
