Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Padilla-Benavides, T.; George Thompson, A.M.; McEvoy, M.M.; Argueello, J.M.
    Mechanism of ATPase-mediated Cu+ export and delivery to periplasmic chaperones: the interaction of Escherichia coli CopA and CusF (2014), J. Biol. Chem., 289, 20492-20501.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
7.2.2.8 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

EC Number Protein Variants Comment Organism
7.2.2.8 D207A/N208A/M209A/M210A site-directed mutagenesis Escherichia coli
7.2.2.8 E287A site-directed mutagenesis Escherichia coli
7.2.2.8 E287C site-directed mutagenesis Escherichia coli
7.2.2.8 K23A/K30A/K31A/H35A/R50A site-directed mutagenesis Escherichia coli
7.2.2.8 M204A site-directed mutagenesis Escherichia coli
7.2.2.8 M204C site-directed mutagenesis Escherichia coli
7.2.2.8 M279A/E280A/H283A site-directed mutagenesis Escherichia coli
7.2.2.8 additional information interaction analysis of recombinant wild-type and mutant enzymes CopA and chaperones CusF, overview Escherichia coli
7.2.2.8 T212A/D214A/N215A/S217A site-directed mutagenesis Escherichia coli
7.2.2.8 W273A/W276A/F277A site-directed mutagenesis Escherichia coli
7.2.2.8 W797A/T800A/T802A site-directed mutagenesis Escherichia coli

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
7.2.2.8 additional information
-
 additional information Cu+-ATPase activity kinetic parameters and binding stoichiometry of recombinant wild-type and mutant enzymes, overview Escherichia coli

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
7.2.2.8 Mg2+ required Escherichia coli

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
7.2.2.8 ATP + H2O + Cu+[side 1] Escherichia coli
-
 ADP + phosphate + Cu+[side 2]
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
7.2.2.8 Escherichia coli
-
 gene copA
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
7.2.2.8 ATP + H2O + Cu+[side 1]
-
 Escherichia coli ADP + phosphate + Cu+[side 2]
-
 ?
7.2.2.8 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

EC Number Synonyms Comment Organism
7.2.2.8 CopA
-
 Escherichia coli
7.2.2.8 Cu+-ATPase
-
 Escherichia coli
7.2.2.8 EcCopA
-
 Escherichia coli

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
7.2.2.8 22
-
 assay at room temperature Escherichia coli

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
7.2.2.8 8
-
 assay at Escherichia coli

General Information

EC Number General Information Comment Organism
7.2.2.8 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
7.2.2.8 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
7.2.2.8 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