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Cytosolic Copper Binding by a Bacterial Storage Protein and Interplay with Copper Efflux.


ABSTRACT: Escherichia coli has a well-characterized copper (Cu) transporting ATPase (CopA) that removes this potentially toxic metal ion from the cytosol. Growth of the strain lacking CopA (?copA) is inhibited above 0.5 mM Cu, whilst a similar effect does not occur in wild type (WT) E. coli until over 2.5 mM Cu. Limited expression of CopA can restore growth to WT levels in ?copA E. coli in the presence of Cu. To study the influence of a bacterial cytosolic Cu storage protein (Csp3) on how E. coli handles Cu, the protein from Bacillus subtilis (BsCsp3) has been expressed in the WT and ?copA strains. BsCsp3 can protect both strains from Cu toxicity, promoting growth at up to ~1.5 and ~3.5 mM Cu, respectively. Higher levels of Csp3 expression are needed to provide resistance to Cu toxicity in ?copA E. coli. At 1.5 mM Cu, BsCsp3 purified from ?copA E. coli binds up to approximately four equivalents of Cu(I) per monomer. A similar number of Cu(I) equivalents can be bound by BsCsp3 purified from WT E. coli also grown at 1.5 mM Cu, a concentration that does not cause toxicity in this strain. Much lower amounts of BsCsp3 are produced in WT E. coli grown in the presence of 3.4 mM Cu, but the protein still counteracts toxicity and is almost half loaded with Cu(I). Csp3s can protect E. coli from Cu toxicity by sequestering cuprous ions in the cytosol. This appears to include an ability to acquire and withhold Cu(I) from the main efflux system in a heterologous host.

SUBMITTER: Lee J 

PROVIDER: S-EPMC6747150 | biostudies-literature | 2019 Aug

REPOSITORIES: biostudies-literature

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Cytosolic Copper Binding by a Bacterial Storage Protein and Interplay with Copper Efflux.

Lee Jaeick J   Dennison Christopher C  

International journal of molecular sciences 20190825 17


<i>Escherichia coli</i> has a well-characterized copper (Cu) transporting ATPase (CopA) that removes this potentially toxic metal ion from the cytosol. Growth of the strain lacking CopA (Δ<i>copA</i>) is inhibited above 0.5 mM Cu, whilst a similar effect does not occur in wild type (WT) <i>E. coli</i> until over 2.5 mM Cu. Limited expression of CopA can restore growth to WT levels in Δ<i>copA</i> <i>E. coli</i> in the presence of Cu. To study the influence of a bacterial cytosolic Cu storage pro  ...[more]

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