Cloned (Comment) | Organism |
---|---|
gene ggt, recombinant overexpression of wild-type and mutant enzymes | Pseudomonas nitroreducens |
Crystallization (Comment) | Organism |
---|---|
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 | Comment | Organism |
---|---|---|
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 |
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 |
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 | Comment | Organism | Structure |
---|---|---|---|
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 | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas nitroreducens | E0D5C2 | - |
- |
Pseudomonas nitroreducens IFO12694 | E0D5C2 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes | Pseudomonas nitroreducens |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
7.1 | - |
about, recombinant wild-type enzyme, pH 10.5, 30°C, transpeptidation activity | Pseudomonas nitroreducens |
15 | - |
about, recombinant enzyme mutant W385T, pH 10.5, 30°C, transpeptidation activity | Pseudomonas nitroreducens |
16 | - |
about, recombinant enzyme mutant W525A, pH 10.5, 30°C, transpeptidation activity | Pseudomonas nitroreducens |
17 | - |
about, recombinant enzyme mutant F417Y, pH 10.5, 30°C, transpeptidation activity | Pseudomonas nitroreducens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
gamma-glutamyl-4-nitroanilide + hydroxamate | - |
Pseudomonas nitroreducens | 4-nitroaniline + gamma-L-glutamylhydroxamate | - |
? | |
gamma-glutamyl-4-nitroanilide + hydroxamate | - |
Pseudomonas nitroreducens IFO12694 | 4-nitroaniline + gamma-L-glutamylhydroxamate | - |
? | |
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 | ? | - |
- |
|
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 | Comment | Organism |
---|---|---|
gamma-glutamyltranspeptidase | - |
Pseudomonas nitroreducens |
GGT | - |
Pseudomonas nitroreducens |
More | see also EC 3.4.19.13 | Pseudomonas nitroreducens |
PnGGT | - |
Pseudomonas nitroreducens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Pseudomonas nitroreducens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
10.5 | - |
assay at | Pseudomonas nitroreducens |
General Information | Comment | Organism |
---|---|---|
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 |