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Literature summary for 2.3.2.2 extracted from

  • Boanca, G.; Sand, A.; Barycki, J.J.
    Uncoupling the enzymatic and autoprocessing activities of Helicobacter pylori gamma-glutamyltranspeptidase (2006), J. Biol. Chem., 281, 19029-19037.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Helicobacter pylori

Protein Variants

Protein Variants Comment Organism
T380A mutant enzyme can not process even after 30 days at 37 °C, no enzymatic activity Helicobacter pylori
T380S mutant enzyme is strongly impaired with respect to processing. Prolonged incubation (more than 30 days) at 37°C leads to complete maturation of the T380S mutant. Vmax for hydrolysis of 5-L-glutamyl-4-nitroanilide is 8.8fold lower than wild-type enzyme. Vmax for transpeptidation of 5-L-glutamyl-4-nitroanilide of Gly-Gly Helicobacter pylori

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0109
-
5-L-glutamyl-4-nitroanilide 37°C, pH 8.0, transpeptidase reaction with Gly-Gly Helicobacter pylori
0.0125
-
5-L-glutamyl-4-nitroanilide 37°C, pH 8.0, hydrolysis Helicobacter pylori
2.5
-
Gly-Gly 37°C, pH 8.0 Helicobacter pylori

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
20000
-
2 * 20000 + 2 * 40000, SDS-PAGE Helicobacter pylori
40000
-
2 * 20000 + 2 * 40000, SDS-PAGE Helicobacter pylori
120000
-
gel filtration Helicobacter pylori

Organism

Organism UniProt Comment Textmining
Helicobacter pylori
-
-
-

Posttranslational Modification

Posttranslational Modification Comment Organism
proteolytic modification the 60000 Da proenzyme is kinetically competent to form the mature 40000 and 20000 Da subunits and exhibits maximal autoprocessing activity at neutral pH. Processing is required for enzymatic activity Helicobacter pylori

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme Helicobacter pylori

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.59
-
hydrolysis of glutathione Helicobacter pylori

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5-L-glutamyl-4-nitroanilide + Gly-Gly
-
Helicobacter pylori 4-nitroaniline + 5-L-glutamyl-Gly-Gly
-
?
5-L-glutamyl-4-nitroanilide + H2O
-
Helicobacter pylori 4-nitroaniline + L-glutamate
-
?

Subunits

Subunits Comment Organism
tetramer 2 * 20000 + 2 * 40000, SDS-PAGE Helicobacter pylori

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5 8 hydrolysis of 5-L-glutamyl-4-nitroanilide Helicobacter pylori

pH Range

pH Minimum pH Maximum Comment Organism
6 9 pH 6.0: about 40% of maximal activity, pH 9.0: about 70% of maximal activity, hydrolysis of 5-L-glutamyl-4-nitroanilide Helicobacter pylori