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Literature summary extracted from

  • Albino, A.; De Angelis, A.; Marco, S.; Severino, V.; Chambery, A.; Di Maro, A.; Desiderio, D.; Raimo, G.; Masullo, M.; De Vendittis, E.
    The cold-adapted gamma-glutamyl-cysteine ligase from the psychrophile Pseudoalteromonas haloplanktis (2014), Biochimie, 104, 50-60 .
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
6.3.2.2 additional information high activity in 100-300 mM Tris buffer Pseudoalteromonas haloplanktis

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.3.2.2 gene gshA, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) Pseudoalteromonas haloplanktis

Inhibitors

EC Number Inhibitors Comment Organism Structure
6.3.2.2 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.3.2.2 additional information
-
 additional information kinetics, recombinant enzyme Pseudoalteromonas haloplanktis
6.3.2.2 0.05
-
 L-cysteine pH 8.0, 15°C, recombinant enzyme Pseudoalteromonas haloplanktis
6.3.2.2 0.093
-
 ATP pH 8.0, 15°C, recombinant enzyme Pseudoalteromonas haloplanktis
6.3.2.2 2.8
-
 L-glutamate pH 8.0, 15°C, recombinant enzyme Pseudoalteromonas haloplanktis

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
6.3.2.2 Mg2+ required, best at 20 mM Pseudoalteromonas haloplanktis

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
6.3.2.2 54500
-
 recombinant enzyme, gel filtration Pseudoalteromonas haloplanktis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6.3.2.2 ATP + L-glutamate + L-cysteine Pseudoalteromonas haloplanktis
-
 ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
 ?
6.3.2.2 ATP + L-glutamate + L-cysteine Pseudoalteromonas haloplanktis TAC 125
-
 ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
6.3.2.2 Pseudoalteromonas haloplanktis Q3IEB7
-
-
6.3.2.2 Pseudoalteromonas haloplanktis TAC 125 Q3IEB7
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.3.2.2 recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Pseudoalteromonas haloplanktis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.3.2.2 ATP + L-glutamate + 2-aminobutyrate 2-aminobutyrate can replace cysteine, although with a lower activity Pseudoalteromonas haloplanktis ADP + phosphate + gamma-L-glutamyl-(2-aminobutyrate)
-
 ?
6.3.2.2 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)
-
 ?
6.3.2.2 ATP + L-glutamate + L-cysteine
-
 Pseudoalteromonas haloplanktis ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
 ?
6.3.2.2 ATP + L-glutamate + L-cysteine
-
 Pseudoalteromonas haloplanktis TAC 125 ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
 ?

Subunits

EC Number Subunits Comment Organism
6.3.2.2 monomer 1 * 58000, SDS-PAGE, 1 * 57663, mass spectrometry and sequence calculation Pseudoalteromonas haloplanktis
6.3.2.2 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

EC Number Synonyms Comment Organism
6.3.2.2 gamma-glutamyl-cysteine ligase
-
 Pseudoalteromonas haloplanktis
6.3.2.2 GshA
-
 Pseudoalteromonas haloplanktis
6.3.2.2 PhGshA II
-
 Pseudoalteromonas haloplanktis
6.3.2.2 PSHAa0937
-
 Pseudoalteromonas haloplanktis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
6.3.2.2 28
-
-
 Pseudoalteromonas haloplanktis

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
6.3.2.2 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
6.3.2.2 10 35 activity range, profile overview Pseudoalteromonas haloplanktis

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
6.3.2.2 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

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
6.3.2.2 8
-
 assay at Pseudoalteromonas haloplanktis

pH Range

EC Number pH Minimum pH Maximum Comment Organism
6.3.2.2 7 9 narrow pH range, highest activity at pH 7.8-8.6 Pseudoalteromonas haloplanktis

Cofactor

EC Number Cofactor Comment Organism Structure
6.3.2.2 ATP
-
 Pseudoalteromonas haloplanktis

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
6.3.2.2 additional information
-
 additional information inhibition kinetics of GSH and GSSG, overview Pseudoalteromonas haloplanktis

General Information

EC Number General Information Comment Organism
6.3.2.2 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
6.3.2.2 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]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
6.3.2.2 0.044
-
 ATP pH 8.0, 15°C, recombinant enzyme Pseudoalteromonas haloplanktis