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Literature summary for 7.1.2.2 extracted from

  • Fujisawa, Y.; Kato, K.; Giulivi, C.
    Nitration of tyrosine residues 368 and 345 in the beta-subunit elicits FoF1-ATPase activity loss (2009), Biochem. J., 423, 219-231.
    View publication on PubMed

Cloned(Commentary)

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

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

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

Metals/Ions Comment Organism Structure
Mg2+ required Rattus norvegicus

Natural Substrates/ Products (Substrates)

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

Organism UniProt Comment Textmining
Rattus norvegicus P10719 beta-subunit; Sprague-Dawley rats
-

Posttranslational Modification

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 (Commentary)

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 (Commentary)

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 [micromol/min/mg]

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 and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ADP + phosphate + H+/out
-
Rattus norvegicus ATP + H2O + H+/in
-
r

Subunits

Subunits Comment Organism
More three-dimensional structures of wild-type and mutant beta-subunits, overview Rattus norvegicus

Synonyms

Synonyms Comment Organism
FoF1-ATPase
-
Rattus norvegicus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
27
-
ATPase activity assay at Rattus norvegicus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
ATPase activity assay at Rattus norvegicus