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Literature summary extracted from

  • Hibi, T.; Imaoka, M.; Shimizu, Y.; Itoh, T.; Wakayama, M.
    Crystal structure analysis and enzymatic characterization of gamma-glutamyltranspeptidase from Pseudomonas nitroreducens (2019), Biosci. Biotechnol. Biochem., 83, 262-269 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.3.2.2 gene ggt, recombinant overexpression of wild-type and mutant enzymes Pseudomonas nitroreducens

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.3.2.2 purified recombinant wild-type PnGGT enzyme, sitting drop vapor diffusion method, mixing of 0.001 ml of 3-5 mg/ml protein in 0.1 M HEPES, pH 7.0, with 0.001 ml reservoir solution containing 16% PEG 8000, 15% PEG 400, 0.1 M HEPES, pH 7.0, 50 mM glycylglycine, and equilibration against 0.5 ml of reservoir solution, 20°C, 1 week, method optimization, X-ray diffraction structure determination and analysis at 1.57-1.70 A resolution, molecular replacement method using the structure of Escherichia coli GGT (EcGGT, PDB ID 2DG5) as the template, modeling Pseudomonas nitroreducens

Protein Variants

EC Number Protein Variants Comment Organism
2.3.2.2 F417Y site-directed mutagenesis, the mutant shows decreased hydrolysis and increased transfer activity, i.e. gamma-glutamyl-p-nitroanilide hydrolysis and gamma-L-glutamylhydroxamate synthesis, compared to wild-type Pseudomonas nitroreducens
2.3.2.2 W385T site-directed mutagenesis, the mutant shows decreased hydrolysis and increased transfer activity, i.e. gamma-glutamyl-p-nitroanilide hydrolysis and gamma-L-glutamylhydroxamate synthesis, compared to wild-type Pseudomonas nitroreducens
2.3.2.2 W525A site-directed mutagenesis, the mutant shows decreased hydrolysis and increased transfer activity, i.e. gamma-glutamyl-p-nitroanilide hydrolysis and gamma-L-glutamylhydroxamate synthesis, compared to wild-type Pseudomonas nitroreducens

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.3.2.2 2-amino-4-([3-(carboxymethyl)phenoxy](methoyl)phosphoryl)butanoic acid peptidyl phosphonate inhibitor GGsTop, the hydroxy-2-oxoethylamino-oxidanylidene-butane moiety of the inhibitor mimicks an acceptor substrate Pseudomonas nitroreducens

Organism

EC Number Organism UniProt Comment Textmining
2.3.2.2 Pseudomonas nitroreducens E0D5C2
-
-
2.3.2.2 Pseudomonas nitroreducens IFO12694 E0D5C2
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.3.2.2 recombinant wild-type and mutant enzymes Pseudomonas nitroreducens

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.3.2.2 7.1
-
 about, recombinant wild-type enzyme, pH 10.5, 30°C, transpeptidation activity Pseudomonas nitroreducens
2.3.2.2 15
-
 about, recombinant enzyme mutant W385T, pH 10.5, 30°C, transpeptidation activity Pseudomonas nitroreducens
2.3.2.2 16
-
 about, recombinant enzyme mutant W525A, pH 10.5, 30°C, transpeptidation activity Pseudomonas nitroreducens
2.3.2.2 17
-
 about, recombinant enzyme mutant F417Y, pH 10.5, 30°C, transpeptidation activity Pseudomonas nitroreducens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.3.2.2 gamma-glutamyl-4-nitroanilide + hydroxamate
-
 Pseudomonas nitroreducens 4-nitroaniline + gamma-L-glutamylhydroxamate
-
 ?
2.3.2.2 gamma-glutamyl-4-nitroanilide + hydroxamate
-
 Pseudomonas nitroreducens IFO12694 4-nitroaniline + gamma-L-glutamylhydroxamate
-
 ?
2.3.2.2 additional information the gamma-glutamyltranspeptidase (GGT) from Pseudomonas nitroreducens strain IFO12694 (PnGGT) has a unique preference for primary amines as gamma-glutamyl acceptors over standard L-amino acids and peptides, detection of the structural basis of PnGGT's hydrolysis and transpeptidation reactions, overview. Key interactions between three residues Trp385, Phe417, and Trp525 distinguish PnGGT from other GGTs, the aromatic side chains of Trp385, Phe417, and Trp525 are involved in the recognition of acceptor substrates. Local structures of substrate binding sites Pseudomonas nitroreducens ?
-
-
2.3.2.2 additional information the gamma-glutamyltranspeptidase (GGT) from Pseudomonas nitroreducens strain IFO12694 (PnGGT) has a unique preference for primary amines as gamma-glutamyl acceptors over standard L-amino acids and peptides, detection of the structural basis of PnGGT's hydrolysis and transpeptidation reactions, overview. Key interactions between three residues Trp385, Phe417, and Trp525 distinguish PnGGT from other GGTs, the aromatic side chains of Trp385, Phe417, and Trp525 are involved in the recognition of acceptor substrates. Local structures of substrate binding sites Pseudomonas nitroreducens IFO12694 ?
-
-

Synonyms

EC Number Synonyms Comment Organism
2.3.2.2 gamma-glutamyltranspeptidase
-
 Pseudomonas nitroreducens
2.3.2.2 GGT
-
 Pseudomonas nitroreducens
2.3.2.2 More see also EC 3.4.19.13 Pseudomonas nitroreducens
2.3.2.2 PnGGT
-
 Pseudomonas nitroreducens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.3.2.2 30
-
 assay at Pseudomonas nitroreducens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.3.2.2 10.5
-
 assay at Pseudomonas nitroreducens

General Information

EC Number General Information Comment Organism
2.3.2.2 additional information comparisons of the active site structures of GGT enzyme from Pseudomonas nitroreducens with those from Escherichia coli and Homo sapiens, overview. The residues around the donor substrate-binding site of PnGGT (Arg94, Asn384, Glu403, Asp406, Ser435, Ser436, Gly456, and Gly456) are conserved among the other GGTs. In the active site of PnGGT, Phe417 in the 411-421 loop and Trp385 form the side wall of the postulated acceptor-binding pocket Pseudomonas nitroreducens