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Literature summary for 7.2.2.8 extracted from

  • Padilla-Benavides, T.; McCann, C.J.; Argüello, J.M.
    The mechanism of Cu+ transport ATPases: interaction with CU+ chaperones and the role of transient metal-binding sites (2012), J. Biol. Chem., 288, 69-78.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
CopA cDNA lacking the N-terminal metal binding domain and the C-terminal metal-binding domain coding regions is ligated into pEXP-NT vector, which adds an N terminus His6 tag sequence is used as a template to introduce the mutations coding for the single substitutions M158A, M158C, E205A, E205C, D336A, and D336C, and the multiple replacements S139A/G140A and K145A/S149A/R152A/R153A/R154A Archaeoglobus fulgidus
expressed in Escherichia coli BL21(DE3) cells Archaeoglobus fulgidus

Protein Variants

Protein Variants Comment Organism
D336A free Cu+/ATPase activity kinetic parameters and binding stoichiometry of wild type and mutant enzyme Archaeoglobus fulgidus
D336A mutation does not affect the ATPase activity when stimulated by free Cu+ in the assay media, nor does the mutation impair Cu+ binding to transmembrane metal-binding site Archaeoglobus fulgidus
D336A the mutant shows increased Vmax value compared to the wild type enzyme Archaeoglobus fulgidus
D336C free Cu+/ATPase activity kinetic parameters and binding stoichiometry of wild type and mutant enzyme Archaeoglobus fulgidus
D336C mutation does not affect the ATPase activity when stimulated by free Cu+ in the assay media, nor does the mutation impair Cu+ binding to transmembrane metal-binding site Archaeoglobus fulgidus
D336C the mutant shows increased Vmax value compared to the wild type enzyme Archaeoglobus fulgidus
E205A free Cu+/ATPase activity kinetic parameters and binding stoichiometry of wild type and mutant enzyme Archaeoglobus fulgidus
E205A mutation does not affect the ATPase activity when stimulated by free Cu+ in the assay media, nor does the mutation impair Cu+ binding to transmembrane metal-binding site Archaeoglobus fulgidus
E205A the mutant shows increased Vmax value compared to the wild type enzyme Archaeoglobus fulgidus
E205C free Cu+/ATPase activity kinetic parameters and binding stoichiometry of wild type and mutant enzyme Archaeoglobus fulgidus
E205C mutation does not affect the ATPase activity when stimulated by free Cu+ in the assay media, nor does the mutation impair Cu+ binding to transmembrane metal-binding site Archaeoglobus fulgidus
E205C the mutant shows increased Vmax value compared to the wild type enzyme Archaeoglobus fulgidus
K145A/S149A/R152A/R153A/R154A mutation does not affect the ATPase activity when stimulated by free Cu+ in the assay media, nor does the mutation impair Cu+ binding to transmembrane metal-binding site Archaeoglobus fulgidus
M158A free Cu+/ATPase activity kinetic parameters and binding stoichiometry of wild type and mutant enzyme Archaeoglobus fulgidus
M158A mutation does not affect the ATPase activity when stimulated by free Cu+ in the assay media, nor does the mutation impair Cu+ binding to transmembrane metal-binding site Archaeoglobus fulgidus
M158A the mutant shows reduced Vmax value compared to the wild type enzyme Archaeoglobus fulgidus
M158C free Cu+/ATPase activity kinetic parameters and binding stoichiometry of wild type and mutant enzyme Archaeoglobus fulgidus
M158C mutation does not affect the ATPase activity when stimulated by free Cu+ in the assay media, nor does the mutation impair Cu+ binding to transmembrane metal-binding site Archaeoglobus fulgidus
M158C the mutant shows increased Vmax value compared to the wild type enzyme Archaeoglobus fulgidus
S139A/G140A free Cu+/ATPase activity kinetic parameters and binding stoichiometry of wild type and mutant enzyme Archaeoglobus fulgidus
S139A/G140A mutation does not affect the ATPase activity when stimulated by free Cu+ in the assay media, nor does the mutation impair Cu+ binding to transmembrane metal-binding site Archaeoglobus fulgidus
S139A/G140A the mutant shows increased Vmax value compared to the wild type enzyme Archaeoglobus fulgidus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information free Cu+/ATPase activity kinetic parameters and binding stoichiometry of wild type, and mutants M158A, M158C, E205A, E205C, D336A, D336C and S139A/G140A Archaeoglobus fulgidus

Localization

Localization Comment Organism GeneOntology No. Textmining
membrane
-
Archaeoglobus fulgidus 16020
-

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + H2O + Cu+[side 1] Archaeoglobus fulgidus
-
ADP + phosphate + Cu+[side 2]
-
?

Organism

Organism UniProt Comment Textmining
Archaeoglobus fulgidus
-
-
-
Archaeoglobus fulgidus O29777
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Archaeoglobus fulgidus
Ni-NTA column chromatography Archaeoglobus fulgidus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + H2O + Cu+[side 1]
-
Archaeoglobus fulgidus ADP + phosphate + Cu+[side 2]
-
?
additional information experimental evidence supports a model where the Cu+-loaded chaperone interacts with an electropositive platform formed by a kink in the second transmembrane helix of the ATPase. Subsequently, three invariant ATPase residues participate in the ligand exchange that mobilizes Cu+ from the chaperone to the transmembrane helices-metal-binding sites Archaeoglobus fulgidus ?
-
?

Synonyms

Synonyms Comment Organism
CopA
-
Archaeoglobus fulgidus
Cu+ transport ATPase
-
Archaeoglobus fulgidus
Cu+-ATPase
-
Archaeoglobus fulgidus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
additional information
-
additional information free Cu+/ATPase activity kinetic parameters and binding stoichiometry pf wild type, and mutants M158A, M158C, E205A, E205C, D336A, D336C and S139A/G140A Archaeoglobus fulgidus

General Information

General Information Comment Organism
physiological function the enzyme couples the hydrolysis of ATP to the efflux of cytoplasmic Cu+ Archaeoglobus fulgidus