Cloned (Comment) | Organism |
---|---|
expression of His-tagged wild-type and mutant enzyme alpha, beta, and gamma subunits in Escherichia coli strain M15 | Rattus norvegicus |
Protein Variants | Comment | Organism |
---|---|---|
betaY345F | site-directed mutagenesis, the mutant shows 72% reduced inactivation by tyrosine nitration compared to the wild-type enzyme | Rattus norvegicus |
betaY345F/Y368F | site-directed mutagenesis, the mutant shows 99% reduced inactivation by tyrosine nitration compared to the wild-type enzyme | Rattus norvegicus |
betaY368F | site-directed mutagenesis, the mutant shows 46% reduced inactivation by tyrosine nitration compared to the wild-type enzyme | Rattus norvegicus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
additional information | tyrosine nitration is a covalent post-translational protein modification associated with various diseases related to oxidative/nitrative stress, that leads to inactivation of the ATPase activity of the enzyme | Rattus norvegicus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Rattus norvegicus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ADP + phosphate + H+/out | Rattus norvegicus | - |
ATP + H2O + H+/in | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rattus norvegicus | P10719 | beta-subunit; Sprague-Dawley rats | - |
Posttranslational Modification | Comment | Organism |
---|---|---|
additional information | tyrosine nitration is a covalent post-translational protein modification associated with various diseases related to oxidative/nitrative stress, that leads to inactivation of the ATPase activity of the enzyme. Mechanism and stoichiometry of the reaction of Tyr residues with tetranitromethane, overview | Rattus norvegicus |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzyme alpha, beta, and gamma subunits from Escherichia coli strain M15 by nickel affinity chromatography | Rattus norvegicus |
Renatured (Comment) | Organism |
---|---|
F1 is reconstituted by using 3:3:1 molar ratios of recombinant alpha-, beta-, and gamma-subunits with a total protein concentration of 0.1 mg/ml, in the reconstitution dissolving buffer containing 10 mM Tris/succinate, pH 6.0, 0.2 mM 2-mercaptoethanol, 10% glycerol and 1% CHAPS, followed by dialysis against the reconstitution dialysis buffer containing 50 mM Tris/succinate, pH 6.0, 0.05 mM deferoxamine mesylate, 5 mM ATP, 2 mM MgCl2, 0.2 mM 2-mercaptoethanol, 10% glycerol and 1% CHAPS under constant stirring at room temperature | Rattus norvegicus |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
activity of nitrated purified reconstituted recombinant wild-type and mutant F1-ATP synthases, overview | Rattus norvegicus |
1.77 | - |
purified reconstituted recombinant mutant Y345F/Y368F F1-ATP synthase, ATPase activity, pH 7.5, 27°C | Rattus norvegicus |
2.017 | - |
purified reconstituted recombinant mutant Y368F F1-ATP synthase, ATPase activity, pH 7.5, 27°C | Rattus norvegicus |
2.149 | - |
purified reconstituted recombinant mutant Y345F F1-ATP synthase, ATPase activity, pH 7.5, 27°C | Rattus norvegicus |
2.402 | - |
purified reconstituted recombinant wild-type F1-ATP synthase, ATPase activity, pH 7.5, 27°C | Rattus norvegicus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ADP + phosphate + H+/out | - |
Rattus norvegicus | ATP + H2O + H+/in | - |
r |
Subunits | Comment | Organism |
---|---|---|
More | three-dimensional structures of wild-type and mutant beta-subunits, overview | Rattus norvegicus |
Synonyms | Comment | Organism |
---|---|---|
FoF1-ATPase | - |
Rattus norvegicus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
27 | - |
ATPase activity assay at | Rattus norvegicus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
ATPase activity assay at | Rattus norvegicus |