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Literature summary for 6.3.2.2 extracted from
- The cold-adapted gamma-glutamyl-cysteine ligase from the psychrophile Pseudoalteromonas haloplanktis (2014), Biochimie, 104, 50-60 .
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| additional information | high activity in 100-300 mM Tris buffer | Pseudoalteromonas haloplanktis |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene gshA, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) | Pseudoalteromonas haloplanktis |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| glutathione | reduced glutathione (GSH) acts as a probably allosteric inhibitor of rPhGshA II. The oxidised form of glutathione (GSSG) inhibits the enzyme with a more complex inhibition profile, due to the complete monoglutathionylation of rPhGshA II on Cys 386, as proved by mass spectrometry data. Inhibition profiles and kinetics of GSH and GSSG, overview | Pseudoalteromonas haloplanktis |  |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | kinetics, recombinant enzyme | Pseudoalteromonas haloplanktis | |
| 0.05 | - |
L-cysteine | pH 8.0, 15°C, recombinant enzyme | Pseudoalteromonas haloplanktis | |
| 0.093 | - |
ATP | pH 8.0, 15°C, recombinant enzyme | Pseudoalteromonas haloplanktis | |
| 2.8 | - |
L-glutamate | pH 8.0, 15°C, recombinant enzyme | Pseudoalteromonas haloplanktis | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Mg2+ | required, best at 20 mM | Pseudoalteromonas haloplanktis | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 54500 | - |
recombinant enzyme, gel filtration | Pseudoalteromonas haloplanktis |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ATP + L-glutamate + L-cysteine | Pseudoalteromonas haloplanktis | - |
ADP + phosphate + gamma-L-glutamyl-L-cysteine | - |
? | |
| ATP + L-glutamate + L-cysteine | Pseudoalteromonas haloplanktis TAC 125 | - |
ADP + phosphate + gamma-L-glutamyl-L-cysteine | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Pseudoalteromonas haloplanktis | Q3IEB7 | - |
- |
| Pseudoalteromonas haloplanktis TAC 125 | Q3IEB7 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Pseudoalteromonas haloplanktis |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ATP + L-glutamate + 2-aminobutyrate | 2-aminobutyrate can replace cysteine, although with a lower activity | Pseudoalteromonas haloplanktis | ADP + phosphate + gamma-L-glutamyl-(2-aminobutyrate) | - |
? | |
| ATP + L-glutamate + 2-aminobutyrate | 2-aminobutyrate can replace cysteine, although with a lower activity | Pseudoalteromonas haloplanktis TAC 125 | ADP + phosphate + gamma-L-glutamyl-(2-aminobutyrate) | - |
? | |
| ATP + L-glutamate + L-cysteine | - |
Pseudoalteromonas haloplanktis | ADP + phosphate + gamma-L-glutamyl-L-cysteine | - |
? | |
| ATP + L-glutamate + L-cysteine | - |
Pseudoalteromonas haloplanktis TAC 125 | ADP + phosphate + gamma-L-glutamyl-L-cysteine | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| monomer | 1 * 58000, SDS-PAGE, 1 * 57663, mass spectrometry and sequence calculation | Pseudoalteromonas haloplanktis |
| More | three-dimensional model of rPhGshA II obtained by homology modelling, overview. The catalytic residue Cys 386 is located at the protein surface | Pseudoalteromonas haloplanktis |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| gamma-glutamyl-cysteine ligase | - |
Pseudoalteromonas haloplanktis |
| GshA | - |
Pseudoalteromonas haloplanktis |
| PhGshA II | - |
Pseudoalteromonas haloplanktis |
| PSHAa0937 | - |
Pseudoalteromonas haloplanktis |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 28 | - |
- |
Pseudoalteromonas haloplanktis |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| additional information | - |
recombinant PhGshA II possesses more typical features of a psychrophilic enzyme, as it is endowed with lower thermodependence and higher heat sensitivity | Pseudoalteromonas haloplanktis |
| 10 | 35 | activity range, profile overview | Pseudoalteromonas haloplanktis |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| additional information | - |
recombinant PhGshA II possesses more typical features of a psychrophilic enzyme, as it is endowed with lower thermodependence and higher heat sensitivity | Pseudoalteromonas haloplanktis |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 8 | - |
assay at | Pseudoalteromonas haloplanktis |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 7 | 9 | narrow pH range, highest activity at pH 7.8-8.6 | Pseudoalteromonas haloplanktis |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| ATP | - |
Pseudoalteromonas haloplanktis | |
Ki Value [mM]
| Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | inhibition kinetics of GSH and GSSG, overview | Pseudoalteromonas haloplanktis |
General Information
| General Information | Comment | Organism |
|---|---|---|
| metabolism | the enzyme catalyses the first step of glutathione biosynthesis by forming gamma-glutamyl-cysteine from glutamate and cysteine in an ATP-dependent reaction. Formation of gamma-glutamyl-cysteine is not the rate-limiting step of glutathione biosynthesis in Pseudoalteromonas haloplanktis | Pseudoalteromonas haloplanktis |
| additional information | MALDI-TOF mass spectrometric analysis of tryptic peptides, mapping of the identify the cysteinyl residue target of the S-glutathionylation reaction, which occurs at the Cys residue at position 386. Three-dimensional model of rPhGshA II obtained by homology modelling, overview. The catalytic residue Cys 386 is located at the protein surface | Pseudoalteromonas haloplanktis |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.044 | - |
ATP | pH 8.0, 15°C, recombinant enzyme | Pseudoalteromonas haloplanktis | |
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Release 2023.1 (January 2023)
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