Activating Compound | Comment | Organism | Structure |
---|---|---|---|
ATP | the enzyme reaches its maximal activity at an ATP concentration of about 1.4 mM. The enzyme activity decreases by 20% when the concentration of ATP is higher than the physiologically relevant concentration (about 5 mM) | Methanocaldococcus jannaschii | |
CTP | the activity of the enzyme reaches its maximal activity at a CTP concentration of about 1.4 mM. The maximal activity decreases by 68 and 95% when the concentration of CTP is 5.7 and 11.4 mM, respectively | Methanocaldococcus jannaschii |
Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21-Codon Plus (DE3)-RIL cells | Methanocaldococcus jannaschii |
Protein Variants | Comment | Organism |
---|---|---|
C126S | the mutation does not reduce the protein's heat stability or solubility, the mutant contains less than 0.8 and less than 0.08 mol of Mg and Fe per protomer. In the presence of MgCl2, the mutant has activity about 2times higher than that of the wild type enzyme. The activity of the mutant in presence of Co2+ is very low | Methanocaldococcus jannaschii |
C143S | the mutation does not reduce the protein's heat stability or solubility, the mutant contains less than 0.8 and less than 0.08 mol of Mg and Fe per protomer. In the presence of MgCl2, the mutant has activity approximately wild type activity. The activity of the mutant in presence of Co2+ is very low | Methanocaldococcus jannaschii |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Methanocaldococcus jannaschii |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.025 | - |
ATP | apparent value, in 35 mM TES (K+) buffer (pH 7.2) containing 14 mM dithiothreitol, 7 mM MgCl+, at 70°C | Methanocaldococcus jannaschii | |
0.063 | - |
FMN | apparent value, in 35 mM TES (K+) buffer (pH 7.2) containing 14 mM dithiothreitol, 7 mM MgCl+, at 70°C | Methanocaldococcus jannaschii | |
0.48 | - |
CTP | apparent value, in 35 mM TES (K+) buffer (pH 7.2) containing 14 mM dithiothreitol, 7 mM MgCl+, at 70°C | Methanocaldococcus jannaschii |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Co2+ | the enzyme requires divalent metals for activity, the best activity being observed with Co2+, where the activity is 4times greater than that with Mg2+ | Methanocaldococcus jannaschii | |
Fe2+ | the enzyme contains 0.2 mol of iron per protomer | Methanocaldococcus jannaschii | |
Mg2+ | low stimulation of activity | Methanocaldococcus jannaschii | |
Mg2+ | the enzyme contains 8.4 mol of magnesium per protomer | Methanocaldococcus jannaschii | |
Mn2+ | the activity with Mn2+ is 4times lower than that with Co2+ | Methanocaldococcus jannaschii | |
additional information | the enzyme activity is not stimulated by Zn2+ and Fe2+ | Methanocaldococcus jannaschii | |
Ni2+ | weak stimulation of activity | Methanocaldococcus jannaschii |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
17000 | - |
2 * 17000, SDS-PAGE | Methanocaldococcus jannaschii |
29000 | - |
gel filtration | Methanocaldococcus jannaschii |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Methanocaldococcus jannaschii | Q58579 | - |
- |
Oxidation Stability | Organism |
---|---|
aerobically isolated enzyme is active only under reducing conditions (with 5.6 mM dithiothreitol) | Methanocaldococcus jannaschii |
Purification (Comment) | Organism |
---|---|
Mono Q column chromatography | Methanocaldococcus jannaschii |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.001 | - |
using GTP as cosubstrate, in 35 mM TES (K+) buffer (pH 7.2) containing 14 mM dithiothreitol, 7 mM MgCl+, at 70°C | Methanocaldococcus jannaschii |
0.004 | - |
using CTP as cosubstrate, in 35 mM TES (K+) buffer (pH 7.2) containing 14 mM dithiothreitol, 7 mM MgCl+, at 70°C | Methanocaldococcus jannaschii |
0.01 | - |
using ATP as cosubstrate, in 35 mM TES (K+) buffer (pH 7.2) containing 14 mM dithiothreitol, 7 mM MgCl+, at 70°C | Methanocaldococcus jannaschii |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + FMN | the enzyme does not catalyze the reverse reaction to produce FMN and ATP from FAD and diphosphate | Methanocaldococcus jannaschii | diphosphate + FAD | - |
ir | |
CTP + FMN | - |
Methanocaldococcus jannaschii | diphosphate + flavin cytidine dinucleotide | - |
? | |
GTP + FMN | weak specific activity | Methanocaldococcus jannaschii | diphosphate + flavin guanidine dinucleotide | - |
? | |
additional information | the enzyme does not function as a glycerol-3-phosphate cytidylyltransferase because it fails to catalyze the formation of glycerol cytidine dinucleotide when incubated with DL-glycerol 3-phosphate and CTP | Methanocaldococcus jannaschii | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 17000, SDS-PAGE | Methanocaldococcus jannaschii |
Synonyms | Comment | Organism |
---|---|---|
MJ1179 | - |
Methanocaldococcus jannaschii |
RibL | - |
Methanocaldococcus jannaschii |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
31 | - |
ATP | in 35 mM TES (K+) buffer (pH 7.2) containing 14 mM dithiothreitol, 7 mM MgCl+, at 70°C | Methanocaldococcus jannaschii |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.006 | - |
CTP | apparent value, in 35 mM TES (K+) buffer (pH 7.2) containing 14 mM dithiothreitol, 7 mM MgCl+, at 70°C | Methanocaldococcus jannaschii | |
0.064 | - |
FMN | apparent value, in 35 mM TES (K+) buffer (pH 7.2) containing 14 mM dithiothreitol, 7 mM MgCl+, at 70°C | Methanocaldococcus jannaschii | |
0.16 | - |
ATP | apparent value, in 35 mM TES (K+) buffer (pH 7.2) containing 14 mM dithiothreitol, 7 mM MgCl+, at 70°C | Methanocaldococcus jannaschii |