Crystallization (Comment) | Organism |
---|---|
molecular dynamics simulations. Key role for Trp74, in the sealing of the hydrophobic channel connecting the two binding sites, as well as for the two Ala602 and Val605 residues, which form a narrow passage whose opening/closing constitutes an essential event in ammonia transfer | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
A602L | enhanced activity compared to the wild type enzyme, the behaviour of the mutant is similar to that of the wild type counterpart during purification demonstrating no significant modification in the overall protein structure | Escherichia coli |
V605L | enhanced activity compared to the wild type enzyme, the behaviour of the mutant is similar to that of the wild type counterpart during purification demonstrating no significant modification in the overall protein structure | Escherichia coli |
W74A | enhanced activity compared to the wild type enzyme, the behaviour of the mutant is similar to that of the wild type counterpart during purification demonstrating no significant modification in the overall protein structure | Escherichia coli |
W74A | efficiency of ammonia transfer is close to zero. No use of ammonia as a substrate | Escherichia coli |
W74F | enhanced activity compared to the wild type enzyme, the behaviour of the mutant is similar to that of the wild type counterpart during purification demonstrating no significant modification in the overall protein structure | Escherichia coli |
W74L | enhanced activity compared to the wild type enzyme, the behaviour of the mutant is similar to that of the wild type counterpart during purification demonstrating no significant modification in the overall protein structure | Escherichia coli |
W74L | decrease in ammonia transfer, 5-7fold increase in the affinity for glutamine in the presence of fructose 6-phosphate | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0064 | - |
L-gamma-glutamyl-p-nitroanilide | mutant W74F, 37°C | Escherichia coli | |
0.04 | - |
L-glutamine | mutant enzyme W74L, in 50 mM potassium phosphate, pH 7.5, 1 mM EDTA, 50 mM KCl, at 37°C | Escherichia coli | |
0.04 | - |
L-glutamine | mutant W74L, 37°C | Escherichia coli | |
0.048 | - |
L-glutamine | mutant enzyme W74F, in 50 mM potassium phosphate, pH 7.5, 1 mM EDTA, 50 mM KCl, at 37°C | Escherichia coli | |
0.048 | - |
L-glutamine | mutant W74F, 37°C | Escherichia coli | |
0.05 | - |
L-gamma-glutamyl-p-nitroanilide | wild-type, 37°C | Escherichia coli | |
0.072 | - |
L-glutamine | mutant enzyme V605L, in 50 mM potassium phosphate, pH 7.5, 1 mM EDTA, 50 mM KCl, at 37°C | Escherichia coli | |
0.072 | - |
L-glutamine | mutant V605L, 37°C | Escherichia coli | |
0.1 | - |
L-glutamine | mutant enzyme A602L, in 50 mM potassium phosphate, pH 7.5, 1 mM EDTA, 50 mM KCl, at 37°C | Escherichia coli | |
0.1 | - |
L-glutamine | mutant A602L, 37°C | Escherichia coli | |
0.1 | - |
L-gamma-glutamyl-p-nitroanilide | mutant W74L, 37°C | Escherichia coli | |
0.2 | - |
L-gamma-glutamyl-p-nitroanilide | mutant W74A, 37°C | Escherichia coli | |
0.27 | - |
L-glutamine | wild-type, 37°C | Escherichia coli | |
0.27 | - |
L-glutamine | wild type enzyme, in 50 mM potassium phosphate, pH 7.5, 1 mM EDTA, 50 mM KCl, at 37°C | Escherichia coli | |
0.29 | - |
L-gamma-glutamyl-p-nitroanilide | mutant V605L, 37°C | Escherichia coli | |
0.36 | - |
D-fructose 6-phosphate | wild-type, 37°C | Escherichia coli | |
0.36 | - |
D-fructose 6-phosphate | wild type enzyme, in 50 mM potassium phosphate, pH 7.5, 1 mM EDTA, 50 mM KCl, at 37°C | Escherichia coli | |
0.36 | - |
L-gamma-glutamyl-p-nitroanilide | mutant A602L, 37°C | Escherichia coli | |
0.38 | - |
L-glutamine | mutant enzyme W74A, in 50 mM potassium phosphate, pH 7.5, 1 mM EDTA, 50 mM KCl, at 37°C | Escherichia coli | |
0.38 | - |
L-glutamine | mutant W74A, 37°C | Escherichia coli | |
0.67 | - |
D-fructose 6-phosphate | mutant enzyme V605L, in 50 mM potassium phosphate, pH 7.5, 1 mM EDTA, 50 mM KCl, at 37°C | Escherichia coli | |
0.67 | - |
D-fructose 6-phosphate | mutant V605L, 37°C | Escherichia coli | |
0.93 | - |
D-fructose 6-phosphate | mutant enzyme W74A, in 50 mM potassium phosphate, pH 7.5, 1 mM EDTA, 50 mM KCl, at 37°C | Escherichia coli | |
0.93 | - |
D-fructose 6-phosphate | mutant W74A, 37°C | Escherichia coli | |
1.11 | - |
D-fructose 6-phosphate | mutant enzyme W74L, in 50 mM potassium phosphate, pH 7.5, 1 mM EDTA, 50 mM KCl, at 37°C | Escherichia coli | |
1.11 | - |
D-fructose 6-phosphate | mutant W74L, 37°C | Escherichia coli | |
1.15 | - |
D-fructose 6-phosphate | mutant enzyme A602L, in 50 mM potassium phosphate, pH 7.5, 1 mM EDTA, 50 mM KCl, at 37°C | Escherichia coli | |
1.15 | - |
D-fructose 6-phosphate | mutant A602L, 37°C | Escherichia coli | |
1.45 | - |
D-fructose 6-phosphate | mutant enzyme W74F, in 50 mM potassium phosphate, pH 7.5, 1 mM EDTA, 50 mM KCl, at 37°C | Escherichia coli | |
1.45 | - |
D-fructose 6-phosphate | mutant W74F, 37°C | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Escherichia coli | P17169 | - |
- |
Purification (Comment) | Organism |
---|---|
Q-Sepharose fast flow column chromatography and Superdex 200 HR 26/60 Hiload gel filtration | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-fructose 6-phosphate | determination of hexose phosphate-isomerizing activity | Escherichia coli | D-glucose 6-phosphate | - |
? | |
L-gamma-glutamyl-p-nitroanilide + H2O | determination of amidohydrolysing activity | Escherichia coli | L-glutamine + p-nitroaniline | - |
? | |
L-glutamine + D-fructose 6-phosphate | - |
Escherichia coli | L-glutamate + D-glucosamine 6-phosphate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
GlmS | - |
Escherichia coli |
glucosamine-6-P synthase | - |
Escherichia coli |
glucosamine-6-phosphate synthase | - |
Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.08 | - |
D-fructose 6-phosphate | mutant W74L, 37°C | Escherichia coli | |
0.083 | - |
D-fructose 6-phosphate | mutant enzyme W74L, in 50 mM potassium phosphate, pH 7.5, 1 mM EDTA, 50 mM KCl, at 37°C | Escherichia coli | |
0.12 | - |
D-fructose 6-phosphate | mutant enzyme W74A, in 50 mM potassium phosphate, pH 7.5, 1 mM EDTA, 50 mM KCl, at 37°C | Escherichia coli | |
0.12 | - |
D-fructose 6-phosphate | mutant W74A, 37°C | Escherichia coli | |
0.13 | - |
L-gamma-glutamyl-p-nitroanilide | mutant V605L, 37°C | Escherichia coli | |
0.15 | - |
D-fructose 6-phosphate | mutant enzyme V605L, in 50 mM potassium phosphate, pH 7.5, 1 mM EDTA, 50 mM KCl, at 37°C | Escherichia coli | |
0.15 | - |
L-gamma-glutamyl-p-nitroanilide | mutant A602L, 37°C | Escherichia coli | |
0.15 | - |
D-fructose 6-phosphate | mutant V605L, 37°C | Escherichia coli | |
0.17 | - |
L-gamma-glutamyl-p-nitroanilide | wild-type, 37°C | Escherichia coli | |
0.22 | - |
D-fructose 6-phosphate | mutant enzyme A602L, in 50 mM potassium phosphate, pH 7.5, 1 mM EDTA, 50 mM KCl, at 37°C | Escherichia coli | |
0.22 | - |
D-fructose 6-phosphate | mutant A602L, 37°C | Escherichia coli | |
0.28 | - |
L-glutamine | mutant enzyme V605L, in 50 mM potassium phosphate, pH 7.5, 1 mM EDTA, 50 mM KCl, at 37°C | Escherichia coli | |
0.28 | - |
L-glutamine | mutant V605L, 37°C | Escherichia coli | |
0.38 | - |
L-glutamine | mutant enzyme A602L, in 50 mM potassium phosphate, pH 7.5, 1 mM EDTA, 50 mM KCl, at 37°C | Escherichia coli | |
0.38 | - |
L-glutamine | mutant A602L, 37°C | Escherichia coli | |
0.5 | - |
L-gamma-glutamyl-p-nitroanilide | mutant W74L, 37°C | Escherichia coli | |
0.52 | - |
D-fructose 6-phosphate | wild type enzyme, in 50 mM potassium phosphate, pH 7.5, 1 mM EDTA, 50 mM KCl, at 37°C | Escherichia coli | |
0.67 | - |
L-gamma-glutamyl-p-nitroanilide | mutant W74F, 37°C | Escherichia coli | |
0.68 | - |
L-gamma-glutamyl-p-nitroanilide | mutant W74A, 37°C | Escherichia coli | |
0.92 | - |
D-fructose 6-phosphate | mutant enzyme W74F, in 50 mM potassium phosphate, pH 7.5, 1 mM EDTA, 50 mM KCl, at 37°C | Escherichia coli | |
0.92 | - |
D-fructose 6-phosphate | mutant W74F, 37°C | Escherichia coli | |
0.97 | - |
L-glutamine | wild type enzyme, in 50 mM potassium phosphate, pH 7.5, 1 mM EDTA, 50 mM KCl, at 37°C | Escherichia coli | |
1.5 | - |
L-glutamine | mutant W74L, 37°C | Escherichia coli | |
1.55 | - |
L-glutamine | mutant enzyme W74L, in 50 mM potassium phosphate, pH 7.5, 1 mM EDTA, 50 mM KCl, at 37°C | Escherichia coli | |
3 | - |
L-glutamine | mutant W74A, 37°C | Escherichia coli | |
3.02 | - |
L-glutamine | mutant enzyme W74A, in 50 mM potassium phosphate, pH 7.5, 1 mM EDTA, 50 mM KCl, at 37°C | Escherichia coli | |
7.17 | - |
L-glutamine | mutant enzyme W74F, in 50 mM potassium phosphate, pH 7.5, 1 mM EDTA, 50 mM KCl, at 37°C | Escherichia coli | |
7.2 | - |
L-glutamine | mutant W74F, 37°C | Escherichia coli | |
14.4 | - |
D-fructose 6-phosphate | wild-type, 37°C | Escherichia coli | |
14.42 | - |
D-fructose 6-phosphate | wild type enzyme, in 50 mM potassium phosphate, pH 7.5, 1 mM EDTA, 50 mM KCl, at 37°C | Escherichia coli | |
17.17 | - |
L-glutamine | wild type enzyme, in 50 mM potassium phosphate, pH 7.5, 1 mM EDTA, 50 mM KCl, at 37°C | Escherichia coli | |
17.2 | - |
L-glutamine | wild-type, 37°C | Escherichia coli |