Cloned (Comment) | Organism |
---|---|
gene gshB, located on chromosome I, expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) | Pseudoalteromonas haloplanktis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
GSSG | the oxidised form of glutathione acts as an irreversible inhibitor of recombinant GshB | Pseudoalteromonas haloplanktis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.17 | - |
ATP | pH 7.8, 10°C, recombinant enzyme, ATPase activity | Pseudoalteromonas haloplanktis | |
0.18 | - |
ATP | pH 7.8, 20°C, recombinant enzyme, ATPase activity | Pseudoalteromonas haloplanktis | |
0.18 | - |
ATP | pH 7.8, 30°C, recombinant enzyme, ATPase activity | Pseudoalteromonas haloplanktis | |
0.2 | 1 | ATP | pH 7.8, 28°C, recombinant enzyme, ATPase activity | Pseudoalteromonas haloplanktis | |
0.25 | - |
gamma-L-glutamyl-L-cysteine | pH 7.8, 30°C, recombinant enzyme | Pseudoalteromonas haloplanktis | |
0.26 | - |
ATP | pH 7.8, 15°C, recombinant enzyme, ATPase activity | Pseudoalteromonas haloplanktis | |
0.26 | - |
ATP | pH 7.8, 30°C, recombinant enzyme, with gamma-L-glutamyl-L-cysteine and glycine | Pseudoalteromonas haloplanktis | |
0.28 | - |
ATP | pH 7.8, 25°C, recombinant enzyme, ATPase activity | Pseudoalteromonas haloplanktis | |
0.75 | - |
glycine | pH 7.8, 30°C, recombinant enzyme | Pseudoalteromonas haloplanktis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | a divalent cation is absolutely required for the activity, whereas monovalent cations are dispensable. Mg2+ is significantly more effective than Mn2+ | Pseudoalteromonas haloplanktis | |
Mn2+ | a divalent cation is absolutely required for the activity, whereas monovalent cations are dispensable. Mg2+ is significantly more effective than Mn2+ | Pseudoalteromonas haloplanktis |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
35888 | - |
2 * 36000, recombinant His-tagged enzyme, SDS-PAGE, 2 * 35888, sequence calculation | Pseudoalteromonas haloplanktis |
36000 | - |
2 * 36000, recombinant His-tagged enzyme, SDS-PAGE, 2 * 35888, sequence calculation | Pseudoalteromonas haloplanktis |
74000 | - |
recombinant His-tagged enzyme, gel filtration | Pseudoalteromonas haloplanktis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + gamma-L-glutamyl-L-cysteine + glycine | Pseudoalteromonas haloplanktis | - |
ADP + phosphate + glutathione | - |
? | |
ATP + gamma-L-glutamyl-L-cysteine + glycine | Pseudoalteromonas haloplanktis TAC 125 | - |
ADP + phosphate + glutathione | - |
? | |
additional information | Pseudoalteromonas haloplanktis | the enzyme forms a disulfide adduct with 2-mercaptoethanol, when purified in the presence of this reducing agent | ? | - |
? | |
additional information | Pseudoalteromonas haloplanktis TAC 125 | the enzyme forms a disulfide adduct with 2-mercaptoethanol, when purified in the presence of this reducing agent | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudoalteromonas haloplanktis | Q3IFA2 | cold-adapted, gene gshB | - |
Pseudoalteromonas haloplanktis TAC 125 | Q3IFA2 | cold-adapted, gene gshB | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Pseudoalteromonas haloplanktis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + gamma-L-glutamyl-L-cysteine + glycine | - |
Pseudoalteromonas haloplanktis | ADP + phosphate + glutathione | - |
? | |
ATP + gamma-L-glutamyl-L-cysteine + glycine | - |
Pseudoalteromonas haloplanktis TAC 125 | ADP + phosphate + glutathione | - |
? | |
additional information | the enzyme forms a disulfide adduct with 2-mercaptoethanol, when purified in the presence of this reducing agent | Pseudoalteromonas haloplanktis | ? | - |
? | |
additional information | assay method development based on [gamma32P]ATP hydrolysis coupled to the GSH synthesis, overview | Pseudoalteromonas haloplanktis | ? | - |
? | |
additional information | the enzyme forms a disulfide adduct with 2-mercaptoethanol, when purified in the presence of this reducing agent | Pseudoalteromonas haloplanktis TAC 125 | ? | - |
? | |
additional information | assay method development based on [gamma32P]ATP hydrolysis coupled to the GSH synthesis, overview | Pseudoalteromonas haloplanktis TAC 125 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 36000, recombinant His-tagged enzyme, SDS-PAGE, 2 * 35888, sequence calculation | Pseudoalteromonas haloplanktis |
Synonyms | Comment | Organism |
---|---|---|
gshB | - |
Pseudoalteromonas haloplanktis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
- |
Pseudoalteromonas haloplanktis |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
10 | 30 | the recombinant enzyme displays a great temperature-dependent increase in its activity with an unusually high value of energy of activation of 75kJ/mol for a psychrophilic enzyme | Pseudoalteromonas haloplanktis |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
- |
30 | purified recombinant enzyme, pH 7.8, 10 min, completely stable | Pseudoalteromonas haloplanktis |
50.5 | - |
purified recombinant enzyme, pH 7.8, 10 min, 50% inactivation | Pseudoalteromonas haloplanktis |
60 | - |
purified recombinant enzyme, pH 7.8, 10 min, inactivation | Pseudoalteromonas haloplanktis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.87 | - |
ATP | pH 7.8, 10°C, recombinant enzyme, ATPase activity | Pseudoalteromonas haloplanktis | |
1.85 | - |
ATP | pH 7.8, 15°C, recombinant enzyme, ATPase activity | Pseudoalteromonas haloplanktis | |
2.42 | - |
ATP | pH 7.8, 20°C, recombinant enzyme, ATPase activity | Pseudoalteromonas haloplanktis | |
4.72 | - |
ATP | pH 7.8, 25°C, recombinant enzyme, ATPase activity | Pseudoalteromonas haloplanktis | |
4.99 | - |
ATP | pH 7.8, 28°C, recombinant enzyme, ATPase activity | Pseudoalteromonas haloplanktis | |
5.32 | - |
ATP | pH 7.8, 30°C, recombinant enzyme, ATPase activity | Pseudoalteromonas haloplanktis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.4 | 8.6 | - |
Pseudoalteromonas haloplanktis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Pseudoalteromonas haloplanktis |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
10.7 | - |
GSSG | pH 7.8, 30°C, recombinant enzyme | Pseudoalteromonas haloplanktis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
5.1 | - |
ATP | pH 7.8, 10°C, recombinant enzyme, ATPase activity | Pseudoalteromonas haloplanktis | |
7.1 | - |
ATP | pH 7.8, 15°C, recombinant enzyme, ATPase activity | Pseudoalteromonas haloplanktis | |
13.4 | - |
ATP | pH 7.8, 20°C, recombinant enzyme, ATPase activity | Pseudoalteromonas haloplanktis | |
16.9 | - |
ATP | pH 7.8, 25°C, recombinant enzyme, ATPase activity | Pseudoalteromonas haloplanktis | |
23.8 | - |
ATP | pH 7.8, 28°C, recombinant enzyme, ATPase activity | Pseudoalteromonas haloplanktis | |
29.6 | - |
ATP | pH 7.8, 30°C, recombinant enzyme, ATPase activity | Pseudoalteromonas haloplanktis |