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Literature summary for 7.4.2.10 extracted from
- Purification and Characterization of an ATPase GsiA from Salmonella enterica (2017), BioMed Res. Int., 2017, 3076091 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene gsiA, recombinant expression of N-terminally His6-MBP-tagged enzyme in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain DH5alpha | Salmonella enterica subsp. enterica serovar Typhimurium |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| periplasm | - |
Salmonella enterica subsp. enterica serovar Typhimurium | - |
- |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Mg2+ | required | Salmonella enterica subsp. enterica serovar Typhimurium |  |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ATP + H2O | Salmonella enterica subsp. enterica serovar Typhimurium | - |
ADP + phosphate | - |
? | |
| ATP + H2O | Salmonella enterica subsp. enterica serovar Typhimurium SGSC1412 | - |
ADP + phosphate | - |
? | |
| ATP + H2O | Salmonella enterica subsp. enterica serovar Typhimurium ATCC 700720 | - |
ADP + phosphate | - |
? | |
| ATP + H2O + glutathione-[glutathione-binding protein][side 1] | Salmonella enterica subsp. enterica serovar Typhimurium | - |
ADP + phosphate + glutathione[side 2] + [glutathione-binding protein][side 1] | - |
? | |
| ATP + H2O + glutathione-[glutathione-binding protein][side 1] | Salmonella enterica subsp. enterica serovar Typhimurium SGSC1412 | - |
ADP + phosphate + glutathione[side 2] + [glutathione-binding protein][side 1] | - |
? | |
| ATP + H2O + glutathione-[glutathione-binding protein][side 1] | Salmonella enterica subsp. enterica serovar Typhimurium ATCC 700720 | - |
ADP + phosphate + glutathione[side 2] + [glutathione-binding protein][side 1] | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Salmonella enterica subsp. enterica serovar Typhimurium | Q8ZQM4 | - |
- |
| Salmonella enterica subsp. enterica serovar Typhimurium ATCC 700720 | Q8ZQM4 | - |
- |
| Salmonella enterica subsp. enterica serovar Typhimurium SGSC1412 | Q8ZQM4 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-MBP-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity and amylose affinity chromatography, cleavage fo the MBP tag by TEV protease, followed by gel filtration | Salmonella enterica subsp. enterica serovar Typhimurium |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ATP + H2O | - |
Salmonella enterica subsp. enterica serovar Typhimurium | ADP + phosphate | - |
? | |
| ATP + H2O | ATPase activity of GsiA is determined by measuring NADH oxidation through recording the decrease of absorbance at 465 nm. The reaction is carried out in 50 mM Tris/HCl pH 8.0, 300 mM NaCl, and 5% v/v glycerol, containing 800 nM GsiA, 5 mM MgCl2, BSA (0.1 mg/ml), lactate dehydrogenase (0.1 units/ml), 4 mM phospho(enol) pyruvic acid, pyruvate kinase (6 units/ml), 0.32 mM beta-nicotinamide adenine dinucleotide, and reduced dipotassium. The reaction is initiated by adding 1 mM ATP | Salmonella enterica subsp. enterica serovar Typhimurium | ADP + phosphate | - |
? | |
| ATP + H2O | - |
Salmonella enterica subsp. enterica serovar Typhimurium SGSC1412 | ADP + phosphate | - |
? | |
| ATP + H2O | ATPase activity of GsiA is determined by measuring NADH oxidation through recording the decrease of absorbance at 465 nm. The reaction is carried out in 50 mM Tris/HCl pH 8.0, 300 mM NaCl, and 5% v/v glycerol, containing 800 nM GsiA, 5 mM MgCl2, BSA (0.1 mg/ml), lactate dehydrogenase (0.1 units/ml), 4 mM phospho(enol) pyruvic acid, pyruvate kinase (6 units/ml), 0.32 mM beta-nicotinamide adenine dinucleotide, and reduced dipotassium. The reaction is initiated by adding 1 mM ATP | Salmonella enterica subsp. enterica serovar Typhimurium SGSC1412 | ADP + phosphate | - |
? | |
| ATP + H2O | - |
Salmonella enterica subsp. enterica serovar Typhimurium ATCC 700720 | ADP + phosphate | - |
? | |
| ATP + H2O | ATPase activity of GsiA is determined by measuring NADH oxidation through recording the decrease of absorbance at 465 nm. The reaction is carried out in 50 mM Tris/HCl pH 8.0, 300 mM NaCl, and 5% v/v glycerol, containing 800 nM GsiA, 5 mM MgCl2, BSA (0.1 mg/ml), lactate dehydrogenase (0.1 units/ml), 4 mM phospho(enol) pyruvic acid, pyruvate kinase (6 units/ml), 0.32 mM beta-nicotinamide adenine dinucleotide, and reduced dipotassium. The reaction is initiated by adding 1 mM ATP | Salmonella enterica subsp. enterica serovar Typhimurium ATCC 700720 | ADP + phosphate | - |
? | |
| ATP + H2O + glutathione-[glutathione-binding protein][side 1] | - |
Salmonella enterica subsp. enterica serovar Typhimurium | ADP + phosphate + glutathione[side 2] + [glutathione-binding protein][side 1] | - |
? | |
| ATP + H2O + glutathione-[glutathione-binding protein][side 1] | - |
Salmonella enterica subsp. enterica serovar Typhimurium SGSC1412 | ADP + phosphate + glutathione[side 2] + [glutathione-binding protein][side 1] | - |
? | |
| ATP + H2O + glutathione-[glutathione-binding protein][side 1] | - |
Salmonella enterica subsp. enterica serovar Typhimurium ATCC 700720 | ADP + phosphate + glutathione[side 2] + [glutathione-binding protein][side 1] | - |
? | |
| additional information | enzyme GsiA can interact with the transmembrane proteins GsiC and GsiD. GsiA does not interact with GsiB | Salmonella enterica subsp. enterica serovar Typhimurium | ? | - |
- |
|
| additional information | enzyme GsiA can interact with the transmembrane proteins GsiC and GsiD. GsiA does not interact with GsiB | Salmonella enterica subsp. enterica serovar Typhimurium SGSC1412 | ? | - |
- |
|
| additional information | enzyme GsiA can interact with the transmembrane proteins GsiC and GsiD. GsiA does not interact with GsiB | Salmonella enterica subsp. enterica serovar Typhimurium ATCC 700720 | ? | - |
- |
|
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| dimer | active form of GsiA | Salmonella enterica subsp. enterica serovar Typhimurium |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| ATPase | - |
Salmonella enterica subsp. enterica serovar Typhimurium |
| GsiA | - |
Salmonella enterica subsp. enterica serovar Typhimurium |
| STM0848 | - |
Salmonella enterica subsp. enterica serovar Typhimurium |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | - |
ATPase activity, recombinant enzyme | Salmonella enterica subsp. enterica serovar Typhimurium |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 5 | 60 | over 20% of maximal activity within this range | Salmonella enterica subsp. enterica serovar Typhimurium |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 20 | - |
purified recombinant enzyme, pH 8.0, loss of 15% activity within 60 min | Salmonella enterica subsp. enterica serovar Typhimurium |
| 30 | - |
purified recombinant enzyme, pH 8.0, loss of 35% activity within 60 min | Salmonella enterica subsp. enterica serovar Typhimurium |
| 40 | - |
purified recombinant enzyme, pH 8.0, loss of 55% activity within 60 min | Salmonella enterica subsp. enterica serovar Typhimurium |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 8 | - |
ATPase activity, recombinant enzyme | Salmonella enterica subsp. enterica serovar Typhimurium |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 6 | 10 | over 60% of maximal activity within this range | Salmonella enterica subsp. enterica serovar Typhimurium |
pH Stability
| pH Stability | pH Stability Maximum | Comment | Organism |
|---|---|---|---|
| 7 | - |
purified recombinant enzyme, 30°C, 72% activity remaining after 20 min | Salmonella enterica subsp. enterica serovar Typhimurium |
| 8 | - |
purified recombinant enzyme, 30°C, 65% activity remaining after 10 min | Salmonella enterica subsp. enterica serovar Typhimurium |
| 8 | - |
purified recombinant enzyme, 30°C, 75% activity remaining after 40 min | Salmonella enterica subsp. enterica serovar Typhimurium |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | the enzyme belongs to the ABC cassette superfamily | Salmonella enterica subsp. enterica serovar Typhimurium |
| metabolism | enzyme GsiA can interact with the transmembrane proteins GsiC and GsiD. GsiA does not interact with GsiB | Salmonella enterica subsp. enterica serovar Typhimurium |
| physiological function | glutathione is the most important antioxidant in cell and plays a plethora of cellular roles. Salmonella enterica can synthesize glutathione. However, there is a glutathione import like system in Salmonella enterica, and GsiA is the ATP binding protein of glutathione importer | Salmonella enterica subsp. enterica serovar Typhimurium |
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