Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 3.5.1.2 extracted from

  • Hashizume, R.; Maki, Y.; Mizutani, K.; Takahashi, N.; Matsubara, H.; Sugita, A.; Sato, K.; Yamaguchi, S.; Mikami, B.
    Crystal structures of protein glutaminase and its pro forms converted into enzyme-substrate complex (2011), J. Biol. Chem., 286, 38691-38702.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

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 (Commentary)

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

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/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-glutamine + H2O Chryseobacterium proteolyticum
-
L-glutamate + NH3
-
?

Organism

Organism UniProt Comment Textmining
Chryseobacterium proteolyticum
-
gene prgA
-

Purification (Commentary)

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

Reaction Comment Organism Reaction ID
L-glutamine + H2O = L-glutamate + NH3 structure-function relationship and catalytic mechanism, detailed overview Chryseobacterium proteolyticum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-glutamine + H2O
-
Chryseobacterium proteolyticum L-glutamate + NH3
-
?