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Literature summary for 2.1.1.45 extracted from
- Purification of thymidylate synthetase from enzyme-poor sources by affinity chromatography (1980), Methods Enzymol., 66, 709-723.
General Stability
| General Stability | Organism |
|---|---|
| enzyme can be kept active for about 2-3 weeks in buffer F in presence of 0.001 mM methotrexate, dialysis against dilute buffer leads to rapid irreversible inactivation as does freeze-drying or simple freezing | Bos taurus |
| freezing and freeze-drying or dialysis causes rapid inactivation | Escherichia coli |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| 5-fluoro-2'-deoxyuridylic acid | - |
Escherichia coli |  |
| 5-fluoro-2'-deoxyuridylic acid | - |
Lacticaseibacillus casei | |
| 5-fluoro-2'-deoxyuridylic acid | - |
Mus musculus | |
| tetrahydromethotrexate | - |
Bos taurus | |
| tetrahydromethotrexate | - |
Escherichia coli | |
| tetrahydromethotrexate | - |
Lacticaseibacillus casei | |
| tetrahydromethotrexate | - |
Mus musculus | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 35000 | - |
2 * 35000, SDS-PAGE | Mus musculus |
| 35000 | - |
2 * 35000, SDS-PAGE | Bos taurus |
| 70000 | - |
- |
Mus musculus |
| 70000 | - |
gel filtration | Escherichia coli |
| 70000 | - |
gel filtration | Bos taurus |
| 70000 | - |
gel filtration | Lacticaseibacillus casei |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 5,10-methylenetetrahydrofolate + dUMP | Mus musculus | - |
dihydrofolate + dTMP | - |
? | |
| 5,10-methylenetetrahydrofolate + dUMP | Escherichia coli | - |
dihydrofolate + dTMP | - |
? | |
| 5,10-methylenetetrahydrofolate + dUMP | Bos taurus | - |
dihydrofolate + dTMP | - |
? | |
| 5,10-methylenetetrahydrofolate + dUMP | Lacticaseibacillus casei | - |
dihydrofolate + dTMP | - |
? | |
| 5,10-methylenetetrahydrofolate + dUMP | Escherichia coli B / ATCC 11303 | - |
dihydrofolate + dTMP | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Bos taurus | - |
calf | - |
| Escherichia coli | - |
- |
- |
| Escherichia coli B / ATCC 11303 | - |
- |
- |
| Lacticaseibacillus casei | - |
- |
- |
| Mus musculus | - |
Gardner lymphosarcoma | - |
| Mus musculus | - |
mouse, sarcoma 180 | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
- |
Lacticaseibacillus casei |
Source Tissue
| Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|
| thymus | - |
Mus musculus | - |
| thymus | - |
Bos taurus | - |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 0.002135 | - |
Gardner lymphosarcoma | Mus musculus |
| 0.003519 | - |
sarcoma 180 | Mus musculus |
| 0.1024 | - |
- |
Bos taurus |
| 0.6778 | - |
- |
Escherichia coli |
Storage Stability
| Storage Stability | Organism |
|---|---|
| -20°C, occasionally loses activity | Mus musculus |
| -50°C-to -20°C, activity of purified enzyme remains unchanged for 2-3 months | Bos taurus |
| 0-10°C enzyme is stable in buffer D for several weeks | Escherichia coli |
| 0-7°C, concentrated enzyme keeps its activity for 14 days | Bos taurus |
| 0°C, enzyme rapidly loses its activity, more than 90% in 2 h | Mus musculus |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| (6R)-N5,N10-methylene-5,6,7,8-tetrahydrofolate + 2'-deoxyuridylate | - |
Lacticaseibacillus casei | 7,8-dihydrofolate + 5-methyl-2'-deoxyuridylate | - |
r | |
| 5,10-methylenetetrahydrofolate + dUMP | - |
Mus musculus | dihydrofolate + dTMP | - |
? | |
| 5,10-methylenetetrahydrofolate + dUMP | - |
Mus musculus | dihydrofolate + dTMP | - |
r | |
| 5,10-methylenetetrahydrofolate + dUMP | - |
Escherichia coli | dihydrofolate + dTMP | - |
? | |
| 5,10-methylenetetrahydrofolate + dUMP | - |
Escherichia coli | dihydrofolate + dTMP | - |
r | |
| 5,10-methylenetetrahydrofolate + dUMP | - |
Bos taurus | dihydrofolate + dTMP | - |
? | |
| 5,10-methylenetetrahydrofolate + dUMP | - |
Bos taurus | dihydrofolate + dTMP | - |
r | |
| 5,10-methylenetetrahydrofolate + dUMP | - |
Lacticaseibacillus casei | dihydrofolate + dTMP | - |
? | |
| 5,10-methylenetetrahydrofolate + dUMP | - |
Lacticaseibacillus casei | dihydrofolate + dTMP | - |
r | |
| 5,10-methylenetetrahydrofolate + dUMP | - |
Escherichia coli B / ATCC 11303 | dihydrofolate + dTMP | - |
? | |
| 5,10-methylenetetrahydrofolate + dUMP | - |
Escherichia coli B / ATCC 11303 | dihydrofolate + dTMP | - |
r | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| dimer | - |
Escherichia coli |
| dimer | - |
Lacticaseibacillus casei |
| dimer | 2 * 35000, SDS-PAGE | Mus musculus |
| dimer | 2 * 35000, SDS-PAGE | Bos taurus |
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