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Literature summary for 3.4.19.9 extracted from

  • Castellano, I.; Merlino, A.; Rossi, M.; La Cara, F.
    Biochemical and structural properties of gamma-glutamyl transpeptidase from Geobacillus thermodenitrificans: An enzyme specialized in hydrolase activity (2010), Biochimie, 92, 464-474.
    View publication on PubMed
Search for more literature for 3.4.19.9

Cloned(Commentary)

Cloned (Comment) Organism
expressed as a fusion protein containing a C-terminal His-tag in Escherichia coli Geobacillus thermodenitrificans
expression in Escherichia coli Geobacillus thermodenitrificans

Protein Variants

Protein Variants Comment Organism
T353A mutation of the N-terminal residue of the 21 kDa subunit, abolishes the post-translational cleavage of the pro-enzyme, but does not completely block the hydrolytic action Geobacillus thermodenitrificans
T353A mutation abolishes the post-translational cleavage of the pro-enzyme, but does not completely block the hydrolytic action Geobacillus thermodenitrificans

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0076
-
 5-L-glutamyl-4-nitroanilide pH 7.8, 52°C Geobacillus thermodenitrificans
0.0076
-
 L-glutamic acid gamma-(4-nitroanilide) Vmax: 0.36 micromol/min/mg Geobacillus thermodenitrificans

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
21000
-
 SDS-PAGE, shorter subunit after post-translational modification Geobacillus thermodenitrificans
40000
-
 SDS-PAGE, shorter subunit after post-translational modification Geobacillus thermodenitrificans
44000
-
 gel filtration Geobacillus thermodenitrificans
61000
-
 SDS-PAGE, precursor homotetrameric protein of 61000 Da per subunit Geobacillus thermodenitrificans
103000
-
 gel filtration Geobacillus thermodenitrificans
222000
-
 gel filtration Geobacillus thermodenitrificans

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
glutathione + H2O Geobacillus thermodenitrificans enzyme hydrolyses the physiological antioxidant glutathione, suggesting an involvement of the enzyme in the cellular defense mechanism against oxidative stress ?
-
 ?
glutathione + H2O Geobacillus thermodenitrificans NG80-2 enzyme hydrolyses the physiological antioxidant glutathione, suggesting an involvement of the enzyme in the cellular defense mechanism against oxidative stress ?
-
 ?

Organism

Organism UniProt Comment Textmining
Geobacillus thermodenitrificans
-
-
-

Posttranslational Modification

Posttranslational Modification Comment Organism
proteolytic modification the enzyme is synthesized as a precursor homotetrameric protein of 60 kDa per subunit, and undergoes an internal post-translational cleavage of the 60 kDa monomer into 40 kDa and 21 kDa shorter subunits, which are then assembled into an active heterotetramer composed of two 40 kDa and two 21 kDa subunits Geobacillus thermodenitrificans
proteolytic modification precursor homotetrameric protein of 61000 Da per subunit undergoes an internal post-translational cleavage into 40 and 21 kDa shorter subunits, which are then assembled into an active heterotetramer composed of two 40 and two 21 kDa subunits Geobacillus thermodenitrificans

Purification (Commentary)

Purification (Comment) Organism
using Ni-NTA chromatography Geobacillus thermodenitrificans

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5-L-glutamyl-4-nitroanilide + H2O
-
 Geobacillus thermodenitrificans L-glutamic acid + 4-nitroaniline
-
 ?
glutathione + H2O
-
 Geobacillus thermodenitrificans ?
-
 ?
glutathione + H2O enzyme hydrolyses the physiological antioxidant glutathione, suggesting an involvement of the enzyme in the cellular defense mechanism against oxidative stress Geobacillus thermodenitrificans ?
-
 ?
glutathione + H2O
-
 Geobacillus thermodenitrificans NG80-2 ?
-
 ?
glutathione + H2O enzyme hydrolyses the physiological antioxidant glutathione, suggesting an involvement of the enzyme in the cellular defense mechanism against oxidative stress Geobacillus thermodenitrificans NG80-2 ?
-
 ?
L-glutamic acid gamma-(4-nitroanilide) + H2O
-
 Geobacillus thermodenitrificans L-glutamic acid + 4-nitroaniline
-
 ?
additional information enzyme does not show any transpeptidase activity Geobacillus thermodenitrificans ?
-
 ?
additional information enzyme does not show any transpeptidase activity Geobacillus thermodenitrificans NG80-2 ?
-
 ?

Subunits

Subunits Comment Organism
heterotetramer 2 * 21000, 2 * 40000, SDS-PAGE Geobacillus thermodenitrificans
homotetramer 4 * 61000 Da, SDS-PAGE Geobacillus thermodenitrificans
tetramer 2 * 40000 + 2 * 21000, SDS-PAGE. The unprocessed enzyme forms an already active homotetramer, whereas the mature enzyme is a fully active compact alpha2beta2-heterotetramer Geobacillus thermodenitrificans

Synonyms

Synonyms Comment Organism
gamma-glutamyl hydrolase
-
 Geobacillus thermodenitrificans

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
52
-
-
 Geobacillus thermodenitrificans

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
45
-
 24 h, 83% residual activity Geobacillus thermodenitrificans
45
-
 pH 7.8, GthGT retains 83% of the activity after 24 h incubation Geobacillus thermodenitrificans

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.8
-
-
 Geobacillus thermodenitrificans