Cloned (Comment) | Organism |
---|---|
gene copA lacking the N-metal-binding-domain-coding regions, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)lambda | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
D207A/N208A/M209A/M210A | site-directed mutagenesis | Escherichia coli |
E287A | site-directed mutagenesis | Escherichia coli |
E287C | site-directed mutagenesis | Escherichia coli |
K23A/K30A/K31A/H35A/R50A | site-directed mutagenesis | Escherichia coli |
M204A | site-directed mutagenesis | Escherichia coli |
M204C | site-directed mutagenesis | Escherichia coli |
M279A/E280A/H283A | site-directed mutagenesis | Escherichia coli |
additional information | interaction analysis of recombinant wild-type and mutant enzymes CopA and chaperones CusF, overview | Escherichia coli |
T212A/D214A/N215A/S217A | site-directed mutagenesis | Escherichia coli |
W273A/W276A/F277A | site-directed mutagenesis | Escherichia coli |
W797A/T800A/T802A | site-directed mutagenesis | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Cu+-ATPase activity kinetic parameters and binding stoichiometry of recombinant wild-type and mutant enzymes, overview | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + H2O + Cu+[side 1] | Escherichia coli | - |
ADP + phosphate + Cu+[side 2] | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
gene copA | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + H2O + Cu+[side 1] | - |
Escherichia coli | ADP + phosphate + Cu+[side 2] | - |
? | |
ATP + H2O + Cu+[side 1] | in the presence of ATP, all Cu+ is released from the ATPase, dependence of metal transfer on ATP hydrolysis | Escherichia coli | ADP + phosphate + Cu+[side 2] | - |
? |
Synonyms | Comment | Organism |
---|---|---|
CopA | - |
Escherichia coli |
Cu+-ATPase | - |
Escherichia coli |
EcCopA | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
22 | - |
assay at room temperature | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Escherichia coli |
General Information | Comment | Organism |
---|---|---|
malfunction | mutation of CopA extracellular loops or the electropositive surface of CusF leads to a decrease in Cu+ transfer efficiency, while mutation of Met and Glu residues proposed to be part of the metal exit site in the ATPase yields enzymes with lower turnover rates, although Cu+ transfer is minimally affected | Escherichia coli |
additional information | homology modeling of EcCopA, docking of the apo-EcCusF, PDB ID 1ZEQ, or the holo-EcCusF, PDB ID 2VB2, structures with the extracellular periplasmic loops of the ATPase | Escherichia coli |
physiological function | mechanism of ATPase-mediated Cu+ export and delivery to periplasmic chaperones, specific transfer occurs after protein-protein recognition and interaction, requirement of multiple homologous transporters and chaperones for specificity in Cu+ delivery to alternative protein targets. Cellular copper homeostasis requires transmembrane transport and compartmental trafficking while maintaining the cell essentially free of uncomplexed Cu2+/+. In bacteria, soluble cytoplasmic and periplasmic chaperones bind and deliver Cu+ to target transporters or metalloenzymes. Transmembrane Cu+-ATPases couple the hydrolysis of ATP to the efflux of cytoplasmic Cu+. Cytosolic Cu+ chaperones (CopZ) interact with a structural platform in Cu+-ATPases (CopA) and deliver copper into the ion permeation path. CusF is a periplasmic Cu+ chaperone that supplies Cu+ to the CusCBA system for efflux to the extracellular milieu. Direct Cu+ transfer from the ATPase CopA to the periplasmic chaperone CusF requiring the specific interaction of the Cu+-bound form of CopA with apo-CusF for subsequent metal transfer upon ATP hydrolysis, the reverse Cu transfer from CusF to CopA is not observed | Escherichia coli |