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Literature summary for 1.5.1.3 extracted from
- Refolding of recombinant Pneumocystis carinii dihydrofolate reductase and characterization of the enzyme (1993), Protein Expr. Purif., 4, 16-23.
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| Cl- | - |
Pneumocystis carinii |  |
| KCl | - |
Rattus norvegicus | |
| KCl | recombinant enzyme, concentration 0.1 M | Pneumocystis carinii | |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| rat-derived gene is overexpressed in Escherichia coli | Pneumocystis carinii |
Crystallization (Commentary)
| Crystallization (Comment) | Organism |
|---|---|
| crystallization of refolded recombinant enzyme as ternary complex with NADPH and various inhibitors | Pneumocystis carinii |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| KCl | recombinant enzyme, concentration above 0.1 M | Pneumocystis carinii | |
| methotrexate | recombinant enzyme | Pneumocystis carinii | |
| methotrexate | - |
Rattus norvegicus | |
| piritrexim | recombinant enzyme | Pneumocystis carinii | |
| pyrimethamine | - |
Pneumocystis carinii | |
| trimethoprim | - |
Pneumocystis carinii | |
| trimetrexate | recombinant enzyme | Pneumocystis carinii | |
| Urea | recombinant enzyme, above 4 M | Pneumocystis carinii | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | buffer and salt conditions significantly alter the Km-values for the substrates | Pneumocystis carinii | |
| 0.0023 | - |
7,8-dihydrofolate | recombinant enzyme, pH 7.0, 0.1 M imidazole | Pneumocystis carinii | |
| 0.003 | - |
NADPH | recombinant enzyme, pH 7.0, 0.1 M imidazole | Pneumocystis carinii | |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| soluble | recombinant enzyme expressed in Escherichia coli | Pneumocystis carinii | - |
- |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 24000 | - |
recombinant enzyme, gel filtration | Pneumocystis carinii |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Pneumocystis carinii | - |
rat-derived | - |
| Rattus norvegicus | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant purified from Escherichia coli | Pneumocystis carinii |
Renatured (Commentary)
| Renatured (Comment) | Organism |
|---|---|
| refolding after solubilisation from inclusion bodies with 4 M guanidium hydrochloride in 0.5% polyethylene glycol 1450, pH 7.0 | Pneumocystis carinii |
Source Tissue
| Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|
| lung | source of Pneumocystis carnii | Rattus norvegicus | - |
| additional information | derived from rat lung | Pneumocystis carinii | - |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 42 | - |
recombinant enzyme, partially purified, after refolding | Pneumocystis carinii |
| 321 | - |
purified recombinant enzyme | Pneumocystis carinii |
Storage Stability
| Storage Stability | Organism |
|---|---|
| -20°C, 50% glycerol, 30% loss of activity after 9 months | Pneumocystis carinii |
| -70°C, 10% glycerol, less than 5% loss of activity after 9 months | Pneumocystis carinii |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 7,8-dihydrofolate + NADPH | - |
Rattus norvegicus | 5,6,7,8-tetrahydrofolate + NADP+ | - |
? | |
| 7,8-dihydrofolate + NADPH | - |
Pneumocystis carinii | 5,6,7,8-tetrahydrofolate + NADP+ | - |
? | |
| folate + NADPH | no activity | Pneumocystis carinii | 5,6,7,8-tetrahydrofolate + NADP+ | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| monomer | - |
Rattus norvegicus |
| monomer | 1 * 24000, recombinant enzyme, SDS-PAGE | Pneumocystis carinii |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | - |
assay at | Pneumocystis carinii |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 108 | - |
7,8-dihydrofolate | recombinant enzyme | Pneumocystis carinii | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.5 | - |
sharp optimum, recombinant enzyme | Pneumocystis carinii |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 6 | 9 | recombinant, refolded enzyme | Pneumocystis carinii |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| NADPH | - |
Rattus norvegicus | |
| NADPH | - |
Pneumocystis carinii | |
Ki Value [mM]
| Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.00000000002 | - |
methotrexate | recombinant enzyme | Pneumocystis carinii | |
| 0.0007 | - |
pyrimethamine | recombinant enzyme | Pneumocystis carinii | |
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