Literature summary for 4.1.3.27 extracted from

List, F.; Bocola, M.; Haeger, M.C.; Sterner, R.
Constitutively active glutaminase variants provide insights into the activation mechanism of anthranilate synthase (2012), Biochemistry, 51, 2812-2818.

Search for more literature for 4.1.3.27

Activating Compound

Activating Compound	Comment	Organism	Structure
additional information	activation mechanism of anthranilate synthase, overview	Thermotoga maritima

Cloned(Commentary)

Cloned (Comment)	Organism
expression of gene trpE in Escherichia coli strain BL21(DE3)RIPL and of wild-type and mutant genes trpGD and trpG in Escherichia coli strain BL21(DE3)Rosetta, all as N- and C-terminally His6-tagged proteins	Thermotoga maritima

Protein Variants

Protein Variants	Comment	Organism
L126G	site-directed mutagenesis of a residue of the glutaminase subunit TrpG from anthranilate synthase, the mutant constitutively hydrolyzes glutamine in the absence of TrpE in contrast to the wild-type enzyme	Thermotoga maritima
L126G/V127Y/T129Y/Y131V	site-directed mutagenesis of anthranilate synthase glutaminase subunit TrpG residues, the mutant constitutively hydrolyzes glutamine in the absence of TrpE in contrast to the wild-type enzyme	Thermotoga maritima
additional information	in contrast to wild-type TrpG, two TrpG variants with single exchanges constitutively hydrolyze glutamine in the absence of TrpE. The introduced amino acid exchanges result in a distance reduction between the active site Cys-His pair, which facilitates the deprotonation of the sulfhydryl group of the catalytic cysteine and thus enables its nucleophilic attack onto the carboxamide group of the glutamine side chain, molecular dynamics simulations, overview	Thermotoga maritima
T129A	site-directed mutagenesis of a residue of the glutaminase subunit TrpG from anthranilate synthase, the mutant shows no activity with glutamine in absence of TrpE like the wild-type enzyme	Thermotoga maritima
T129F	site-directed mutagenesis of a residue of the glutaminase subunit TrpG from anthranilate synthase, the mutant constitutively hydrolyzes glutamine in the absence of TrpE in contrast to the wild-type enzyme	Thermotoga maritima
T129Y	site-directed mutagenesis of a residue of the glutaminase subunit TrpG from anthranilate synthase, the mutant constitutively hydrolyzes glutamine in the absence of TrpE in contrast to the wild-type enzyme	Thermotoga maritima
V127Y	site-directed mutagenesis of a residue of the glutaminase subunit TrpG from anthranilate synthase, the mutant shows no activity with glutamine in absence of TrpE like the wild-type enzyme	Thermotoga maritima
Y131V	site-directed mutagenesis of a residue of the glutaminase subunit TrpG from anthranilate synthase, the mutant shows no activity with glutamine in absence of TrpE like the wild-type enzyme	Thermotoga maritima

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
additional information	-	additional information	steady-state enzyme kinetics, overview	Thermotoga maritima

Organism

Organism	UniProt	Comment	Textmining
Thermotoga maritima	-	genes trpE, trpGD, and trpG	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant wild-type and mutant His-tagged enzymes from Escherichia coli by nickel affinity and anion exchange chromatography	Thermotoga maritima

Subunits

Subunits	Comment	Organism
More	synthase subunit TrpE and glutaminase subunit TrpG	Thermotoga maritima

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
25	-	assay at	Thermotoga maritima

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8	-	assay at	Thermotoga maritima

General Information

General Information	Comment	Organism
additional information	the enzyme shows a mechanism of this tight activity regulation, catalytic Cys-His-Glu triad, molecular dynamics simulations, overview	Thermotoga maritima

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Release 2023.1 (January 2023)