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Literature summary for 2.7.6.1 extracted from
- Regulation and mechanism of phosphoribosylpyrophosphate synthetase. I. Purification and properties of the enzyme from Salmonella typhimurium (1969), J. Biol. Chem., 244, 2854-2863.
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| 1-Aminoethylphosphonate | 0.1 M, 13% of the activity with phosphate | Salmonella enterica subsp. enterica serovar Typhimurium |  |
| arsenate | 0.1 M, 43% of the activity with phosphate | Salmonella enterica subsp. enterica serovar Typhimurium | |
| phosphate | specific and absolute requirement | Salmonella enterica subsp. enterica serovar Typhimurium | |
| phosphate | appears to have an effect on the stability of the enzyme as well as a specific catalytic effect | Salmonella enterica subsp. enterica serovar Typhimurium | |
| phosphate | the enzyme has 2 sites for phosphate, one of high affinity and one of lower affinity | Salmonella enterica subsp. enterica serovar Typhimurium | |
General Stability
| General Stability | Organism |
|---|---|
| completely inactivated when phosphate is removed by dialysis | Salmonella enterica subsp. enterica serovar Typhimurium |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| ATP | free form, slight inhibition at 1 mM | Salmonella enterica subsp. enterica serovar Typhimurium | |
| GSH | marked inhibition of enzyme in crude extract at 1 mM | Salmonella enterica subsp. enterica serovar Typhimurium | |
| additional information | GSH, 2-mercaptoethanol, 2,3-dimercaptopropanol, dithiothreitol, at 1 mM, have no effect or slightly inhibit the highly purified enzyme | Salmonella enterica subsp. enterica serovar Typhimurium |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.05 | 0.13 | D-ribose 5-phosphate | pH 7.5, 37ºC | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 0.06 | - |
ATP | pH 7.5, 37ºC, excess of Mn2+ | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 0.06 | - |
ATP | ATP in form of MnATP2- | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 0.1 | - |
ATP | pH 7.5, 37ºC, excess of Mg2+, bimodal kinetic behavior | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 0.1 | - |
ATP | ATP in form of MgATP2- | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 0.15 | 0.4 | AMP | pH 7.5, 37ºC | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 0.35 | - |
ATP | pH 7.5, 37ºC, no excess of Mn2+ | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 0.35 | - |
ATP | ATP in form of MnATP2- | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 0.58 | - |
ATP | pH 7.5, 37ºC, excess of Mg2+, bimodal kinetic behavior | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 0.58 | - |
ATP | ATP in form of MgATP2- | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 2.3 | - |
phosphate | pH 7.5, 37ºC, high affinity site | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 2.9 | - |
ATP | pH 7.5, 37ºC, no excess of Mg2+ | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 2.9 | - |
ATP | ATP in form of MgATP2- | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 40 | - |
phosphate | pH 7.5, 37ºC, low affinity site | Salmonella enterica subsp. enterica serovar Typhimurium | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Co2+ | no activity observed | Salmonella enterica subsp. enterica serovar Typhimurium | |
| Mg2+ | requirement | Salmonella enterica subsp. enterica serovar Typhimurium | |
| Mg2+ | required to form a complex with ATP and as a free cation | Salmonella enterica subsp. enterica serovar Typhimurium | |
| Mn2+ | equally effective as Mg2+ | Salmonella enterica subsp. enterica serovar Typhimurium | |
| additional information | absolute requirement for a divalent cation for activity | Salmonella enterica subsp. enterica serovar Typhimurium | |
| additional information | no activity observed with Ca2+ | Salmonella enterica subsp. enterica serovar Typhimurium | |
| Zn2+ | 25% of the activity with Mg2+ | Salmonella enterica subsp. enterica serovar Typhimurium | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 540000 | - |
sucrose density gradient sedimentation | Salmonella enterica subsp. enterica serovar Typhimurium |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ATP + D-ribose 5-phosphate | Salmonella enterica subsp. enterica serovar Typhimurium | the product phosphoribosyldiphosphate is required for the biosynthesis of purine, pyrimidine and pyridine nucleotides, L-histidine and L-tryptophan | AMP + 5-phospho-alpha-D-ribose 1-diphosphate | - |
r | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Salmonella enterica subsp. enterica serovar Typhimurium | - |
LT-2 | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| extraction, streptomycin-heat treatment, ammonium sulfate and acid precipitation, 450fold purification | Salmonella enterica subsp. enterica serovar Typhimurium |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| additional information | - |
26.8 U/mg protein, purified enzyme, 400fold to 450fold the activity of crude extract | Salmonella enterica subsp. enterica serovar Typhimurium |
Storage Stability
| Storage Stability | Organism |
|---|---|
| 0°C or -10°C, 50 mM potassium phosphate buffer, pH 7.4, stable for weeks | Salmonella enterica subsp. enterica serovar Typhimurium |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ATP + D-ribose 5-phosphate | - |
Salmonella enterica subsp. enterica serovar Typhimurium | AMP + 5-phospho-alpha-D-ribose 1-diphosphate | - |
r | |
| ATP + D-ribose 5-phosphate | the product phosphoribosyldiphosphate is required for the biosynthesis of purine, pyrimidine and pyridine nucleotides, L-histidine and L-tryptophan | Salmonella enterica subsp. enterica serovar Typhimurium | AMP + 5-phospho-alpha-D-ribose 1-diphosphate | - |
r | |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | - |
assay at | Salmonella enterica subsp. enterica serovar Typhimurium |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 8.1 | 8.6 | for either the Mg2+ or the Mn2+ supported reaction | Salmonella enterica subsp. enterica serovar Typhimurium |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 6 | 9.5 | very low activity below and above | Salmonella enterica subsp. enterica serovar Typhimurium |
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