EC Number | Cloned (Comment) | Organism |
---|---|---|
7.2.2.8 | 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 |
7.2.2.8 | expressed in Escherichia coli BL21(DE3) cells | Archaeoglobus fulgidus |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
7.2.2.8 | D336A | free Cu+/ATPase activity kinetic parameters and binding stoichiometry of wild type and mutant enzyme | Archaeoglobus fulgidus |
7.2.2.8 | 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 |
7.2.2.8 | D336A | the mutant shows increased Vmax value compared to the wild type enzyme | Archaeoglobus fulgidus |
7.2.2.8 | D336C | free Cu+/ATPase activity kinetic parameters and binding stoichiometry of wild type and mutant enzyme | Archaeoglobus fulgidus |
7.2.2.8 | 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 |
7.2.2.8 | D336C | the mutant shows increased Vmax value compared to the wild type enzyme | Archaeoglobus fulgidus |
7.2.2.8 | E205A | free Cu+/ATPase activity kinetic parameters and binding stoichiometry of wild type and mutant enzyme | Archaeoglobus fulgidus |
7.2.2.8 | 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 |
7.2.2.8 | E205A | the mutant shows increased Vmax value compared to the wild type enzyme | Archaeoglobus fulgidus |
7.2.2.8 | E205C | free Cu+/ATPase activity kinetic parameters and binding stoichiometry of wild type and mutant enzyme | Archaeoglobus fulgidus |
7.2.2.8 | 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 |
7.2.2.8 | E205C | the mutant shows increased Vmax value compared to the wild type enzyme | Archaeoglobus fulgidus |
7.2.2.8 | 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 |
7.2.2.8 | M158A | free Cu+/ATPase activity kinetic parameters and binding stoichiometry of wild type and mutant enzyme | Archaeoglobus fulgidus |
7.2.2.8 | 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 |
7.2.2.8 | M158A | the mutant shows reduced Vmax value compared to the wild type enzyme | Archaeoglobus fulgidus |
7.2.2.8 | M158C | free Cu+/ATPase activity kinetic parameters and binding stoichiometry of wild type and mutant enzyme | Archaeoglobus fulgidus |
7.2.2.8 | 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 |
7.2.2.8 | M158C | the mutant shows increased Vmax value compared to the wild type enzyme | Archaeoglobus fulgidus |
7.2.2.8 | S139A/G140A | free Cu+/ATPase activity kinetic parameters and binding stoichiometry of wild type and mutant enzyme | Archaeoglobus fulgidus |
7.2.2.8 | 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 |
7.2.2.8 | S139A/G140A | the mutant shows increased Vmax value compared to the wild type enzyme | Archaeoglobus fulgidus |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
7.2.2.8 | 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 |
EC Number | Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|---|
7.2.2.8 | membrane | - |
Archaeoglobus fulgidus | 16020 | - |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
7.2.2.8 | ATP + H2O + Cu+[side 1] | Archaeoglobus fulgidus | - |
ADP + phosphate + Cu+[side 2] | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
7.2.2.8 | Archaeoglobus fulgidus | - |
- |
- |
7.2.2.8 | Archaeoglobus fulgidus | O29777 | - |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
7.2.2.8 | - |
Archaeoglobus fulgidus |
7.2.2.8 | Ni-NTA column chromatography | Archaeoglobus fulgidus |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
7.2.2.8 | ATP + H2O + Cu+[side 1] | - |
Archaeoglobus fulgidus | ADP + phosphate + Cu+[side 2] | - |
? | |
7.2.2.8 | 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 | ? | - |
? |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
7.2.2.8 | CopA | - |
Archaeoglobus fulgidus |
7.2.2.8 | Cu+ transport ATPase | - |
Archaeoglobus fulgidus |
7.2.2.8 | Cu+-ATPase | - |
Archaeoglobus fulgidus |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
7.2.2.8 | 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 |
EC Number | General Information | Comment | Organism |
---|---|---|---|
7.2.2.8 | physiological function | the enzyme couples the hydrolysis of ATP to the efflux of cytoplasmic Cu+ | Archaeoglobus fulgidus |