Copper Acetate Manufacturers (WSDTY) proposed synergistic effect in a copper homeostasis-deficient E. coli strain.Based on our expression results, we speculated that Fe(II) might interfere with one or more proteins in the copper homeostasis systems. To test this prediction, we used E. coli as a model organism, because the copper detoxification system is well understood, and Cus (an RND efflux pump), this strain is much more sensitive to copper than its wild-type parent. We grew the parent strain with 1 μM Cu(II) with or without 200 μM Fe(II) and the mutant strain with 300 nM Cu(II) with or without 200 μM Fe(II). The different concentrations of Cu(II) reflected the different copper sensitivities of the parent and mutant strains: we chose Cu(II) concentrations for which each strain showed a reproducible response. We found that when the parent and mutant strains were assayed side by side, the mutant was less affected by Cu(II) with Fe(II) than the wild type: while Cu(II) caused a longer growth lag for the mutant than for the wild type, the addition of Fe(II) increased the growth delay only 2.8-fold for the mutant strain but 23-fold for the parent strain.
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