Cloned (Comment) | Organism |
---|---|
gene prgA, overexpression of C-terminally His6-tagged wild-type and mutant A47Q enzymes in Escherichia coli strain BL21(DE3) cytosol | Chryseobacterium proteolyticum |
Crystallization (Comment) | Organism |
---|---|
purified recombinant mature C-terminally His6-tagged wild-type enzyme, hanging drop vapor diffusion method, mixing 0.005 ml of 34 mg/ml protein in 20 mM NaHEPES, pH 7.0, with 0.005 ml of reservoir solution containing 1.2 M ammonium dihydrogen phosphate, and 0.1 M sodium citrate, pH 5.6, crystallization of wild-type pro-enzyme by sitting drop vapour diffusion, mixing 0.002 ml of 20 mg/ml pro-enzyme in 20 mM NaHEPES, pH 6.5, with 0.002 ml of reservoir solution containing 0.2 M ammonium citrate, pH 5.1, and 20% w/v PEG-3350, crystallization of mutant A47Q-1 by hanging drop vapour diffusion method, by mixing of 0.005 ml of 20 mg/ml protein in 20 mM ammonium tartrate, pH 6.7, with 0.005 ml of reservoir solution containing 0.2 M ammonium tartrate, pH 6.7, and 20% PEG 3350, and of mutant A47Q-2 by mixing 0.005 ml of 20 mg/ml protein in 20 mM sodium phosphate, pH 6.0, with 0.005 ml reservoir solution containing 0.2 M sodium tartrate, pH 8.6, and 20% w/v PEG 3350, 20°C, X-ray diffraction structure determination and analysis at 1.5-1.75 A resolution | Chryseobacterium proteolyticum |
Protein Variants | Comment | Organism |
---|---|---|
A47Q | site-directed mutagenesis, structure analysis, mutant crystal foms A47Q-1 and A47Q-2 at pH 6.7 and pH 8.6, respectively, overview | Chryseobacterium proteolyticum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-glutamine + H2O | Chryseobacterium proteolyticum | - |
L-glutamate + NH3 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Chryseobacterium proteolyticum | - |
gene prgA | - |
Purification (Comment) | Organism |
---|---|
recombinant C-terminally His6-tagged wild-type and mutant A47Q enzymes from Escherichia coli strain BL21(DE3) cytosol by nickel affinity chromatography | Chryseobacterium proteolyticum |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
L-glutamine + H2O = L-glutamate + NH3 | structure-function relationship and catalytic mechanism, detailed overview | Chryseobacterium proteolyticum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-glutamine + H2O | - |
Chryseobacterium proteolyticum | L-glutamate + NH3 | - |
? |