Activating Compound | Comment | Organism | Structure |
---|---|---|---|
phosphate | the enzyme is dependent on phosphate for its activity. The optimal concentration ranges from 10 mM to 40 mM | Mycobacterium tuberculosis | |
SO42- | sulfate ions are able to stimulate the enzyme activity, but with respect to phosphate, are less effective. The optimal concentration ranges from 40 mM to 60 mM | Mycobacterium tuberculosis |
Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3)pLysS cells | Mycobacterium tuberculosis |
General Stability | Organism |
---|---|
dialysis against buffers such as 50 mM Tris-HCl, pH 8.0 or 50 mM HEPES-NaOH, pH 8.0 without phosphate results in a protein precipitation and complete loss of activity. The addition of 50 mM ammonium sulfate or 5 mM Mg-ATP to Tris-HCl, pH 8.0 allows the enzyme to preserve 20% of initial activity after a period of 16 h, whereas full activity is maintained with the addition of 50 mM phosphate | Mycobacterium tuberculosis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
ADP | strong inhibition | Mycobacterium tuberculosis | |
Ca2+ | - |
Mycobacterium tuberculosis | |
Cu2+ | - |
Mycobacterium tuberculosis | |
Fe2+ | - |
Mycobacterium tuberculosis | |
GDP | the enzyme is weakly sensitive to GDP | Mycobacterium tuberculosis | |
additional information | no inhibitory effects are shown by the presence of pyrimidine nucleoside mono- or diphosphates or of histidine, up to 2 mM | Mycobacterium tuberculosis | |
phosphate | - |
Mycobacterium tuberculosis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.06 | - |
D-ribose 5-phosphate | in the presence of 5 mM Mn2+, in 50 mM potassium phosphate pH 8.0, at 37°C | Mycobacterium tuberculosis | |
0.07 | - |
D-ribose 5-phosphate | in the presence of 5 mM Mg2+, in 50 mM potassium phosphate pH 8.0, at 37°C | Mycobacterium tuberculosis | |
0.071 | - |
D-ribose 5-phosphate | in the absence of Mg2+ or Mn2+, in 50 mM potassium phosphate pH 8.0, at 37°C | Mycobacterium tuberculosis |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | allosterically activated by Mg2+ | Mycobacterium tuberculosis | |
Mn2+ | allosterically activated by Mn2+ | Mycobacterium tuberculosis |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
35000 | - |
6 * 35000, SDS-PAGE | Mycobacterium tuberculosis |
220000 | - |
gel filtration | Mycobacterium tuberculosis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mycobacterium tuberculosis | - |
- |
- |
Mycobacterium tuberculosis Rv1017 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
His-Trap HP column chromatography and Superdex 200 gel filtration | Mycobacterium tuberculosis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
Mg-ATP2- + D-ribose 5-phosphate | - |
Mycobacterium tuberculosis | Mg-AMP2- + 5-phospho-alpha-D-ribose 1-diphosphate | - |
? | |
Mg-ATP2- + D-ribose 5-phosphate | - |
Mycobacterium tuberculosis Rv1017 | Mg-AMP2- + 5-phospho-alpha-D-ribose 1-diphosphate | - |
? | |
Mn-ATP2- + D-ribose 5-phosphate | - |
Mycobacterium tuberculosis | Mn-AMP + 5-phospho-alpha-D-ribose 1-diphosphate | - |
? | |
Mn-ATP2- + D-ribose 5-phosphate | - |
Mycobacterium tuberculosis Rv1017 | Mn-AMP + 5-phospho-alpha-D-ribose 1-diphosphate | - |
? |
Subunits | Comment | Organism |
---|---|---|
homohexamer | 6 * 35000, SDS-PAGE | Mycobacterium tuberculosis |
Synonyms | Comment | Organism |
---|---|---|
phosphoribosylpyrophosphate synthetase | - |
Mycobacterium tuberculosis |
PRPPase | - |
Mycobacterium tuberculosis |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
62 | 74 | a highly stable enzyme, losing 50% of its activity in 10 min of incubation at 62°C, and showing a Tm of 69.3°C (without ligands) to 74.5°C (in the presence of 5 mM Mg-ATP) | Mycobacterium tuberculosis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
34.6 | - |
Mg-ATP2- | in the presence of 5 mM Mg2+, in 50 mM potassium phosphate pH 8.0, at 37°C | Mycobacterium tuberculosis | |
35.1 | - |
D-ribose 5-phosphate | in the presence of 5 mM Mg2+, in 50 mM potassium phosphate pH 8.0, at 37°C | Mycobacterium tuberculosis | |
35.5 | - |
Mg-ATP2- | in the absence of free Mg2+ or Mn2+, in 50 mM potassium phosphate pH 8.0, at 37°C | Mycobacterium tuberculosis | |
37 | - |
D-ribose 5-phosphate | in the absence of Mg2+ or Mn2+, in 50 mM potassium phosphate pH 8.0, at 37°C | Mycobacterium tuberculosis | |
44.3 | - |
Mg-ATP2- | in the presence of 5 mM Mn2+, in 50 mM potassium phosphate pH 8.0, at 37°C | Mycobacterium tuberculosis | |
44.7 | - |
D-ribose 5-phosphate | in the presence of 5 mM Mn2+, in 50 mM potassium phosphate pH 8.0, at 37°C | Mycobacterium tuberculosis | |
45.1 | - |
Mn-ATP2- | in the presence of 5 mM Mn2+, in 50 mM potassium phosphate pH 8.0, at 37°C | Mycobacterium tuberculosis | |
46.3 | - |
Mn-ATP2- | in the absence of free Mg2+ or Mn2+, in 50 mM potassium phosphate pH 8.0, at 37°C | Mycobacterium tuberculosis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
the enzyme shows an optimum at a pH value close to 8.0 | Mycobacterium tuberculosis |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
7 | 9.5 | the enzyme possesses nearly 70% of its maximal activity at pH 9.5. The activity at pH 7 is 57% of the maximal one at pH 8.0 | Mycobacterium tuberculosis |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
0.4 | - |
in 50 mM potassium phosphate pH 8.0, at 37°C | Mycobacterium tuberculosis | GDP | |
5 | - |
in 50 mM potassium phosphate pH 8.0, at 37°C | Mycobacterium tuberculosis | GDP | |
100 | - |
in 50 mM potassium phosphate pH 8.0, at 37°C | Mycobacterium tuberculosis | phosphate |
General Information | Comment | Organism |
---|---|---|
physiological function | the enzyme is crucial for mycobacterial cell wall biosynthesis | Mycobacterium tuberculosis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
501.4 | - |
D-ribose 5-phosphate | in the presence of 5 mM Mg2+, in 50 mM potassium phosphate pH 8.0, at 37°C | Mycobacterium tuberculosis | |
521 | - |
D-ribose 5-phosphate | in the absence of Mg2+ or Mn2+, in 50 mM potassium phosphate pH 8.0, at 37°C | Mycobacterium tuberculosis | |
745 | - |
D-ribose 5-phosphate | in the presence of 5 mM Mn2+, in 50 mM potassium phosphate pH 8.0, at 37°C | Mycobacterium tuberculosis |