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Literature summary extracted from
- Effect of dimerization on dihydrofolate reductase catalysis (2013), Biochemistry, 52, 3881-3887 .
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 1.5.1.3 | a variant (Xet-3) of dihydrofolate reductase from Escherichia coli constructed by introducing residues located at the Thermotoga maritima dihydrofolate reductase (TmDHFR) dimer interface is expressed in Escherichia coli BL21(DE3) cells | Escherichia coli |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 1.5.1.3 | additional information | preparation of a dimeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli by introducing residues located at the Thermotoga maritima dihydrofolate reductase (TmDHFR) dimer interface increases the melting temperature to about 60°C, approximately 9°C higher than that measured for the Escherichia coli enzyme. The steady-state and pre-steady-state rate constants measured for Xet-3 were similar to those of dimeric TmDHFR but significantly lower than those of the parent dihydrofolate reductase from Escherichia coli | Escherichia coli |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.5.1.3 | 0.0007 | - |
7,8-dihydrofolate | pH 7, 20°C, wild-type enzyme | Escherichia coli |  |
| 1.5.1.3 | 0.0027 | - |
7,8-dihydrofolate | pH 7, 20°C, monomeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli prepared by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface | Escherichia coli | |
| 1.5.1.3 | 0.0048 | - |
NADPH | pH 7, 20°C, wild-type enzyme | Escherichia coli | |
| 1.5.1.3 | 0.0077 | - |
7,8-dihydrofolate | pH 7, 20°C, dimeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli prepared by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface | Escherichia coli | |
| 1.5.1.3 | 0.0195 | - |
NADPH | pH 7, 20°C, monomeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli prepared by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface | Escherichia coli | |
| 1.5.1.3 | 0.0199 | - |
NADPH | pH 7, 20°C, dimeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli prepared by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface | Escherichia coli | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 1.5.1.3 | Escherichia coli | - |
- |
- |
| 1.5.1.3 | Thermotoga maritima | Q60034 | - |
- |
| 1.5.1.3 | Thermotoga maritima DSM 3109 | Q60034 | - |
- |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 1.5.1.3 | - |
Escherichia coli |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.5.1.3 | 7,8-dihydrofolate + NADPH + H+ | - |
Escherichia coli | 5,6,7,8-tetrahydrofolate + NADP+ | - |
? | |
| 1.5.1.3 | 7,8-dihydrofolate + NADPH + H+ | - |
Thermotoga maritima | 5,6,7,8-tetrahydrofolate + NADP+ | - |
? | |
| 1.5.1.3 | 7,8-dihydrofolate + NADPH + H+ | - |
Thermotoga maritima DSM 3109 | 5,6,7,8-tetrahydrofolate + NADP+ | - |
? | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 1.5.1.3 | homodimer | - |
Thermotoga maritima |
| 1.5.1.3 | monomer | preparation of a dimeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface | Escherichia coli |
Temperature Stability [°C]
| EC Number | Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 1.5.1.3 | 60 | - |
preparation of a dimeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface increases the melting temperature to about 60°C, approximately 9°C higher than that measured for the Escherichia coli enzyme | Escherichia coli |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.5.1.3 | 0.003 | - |
7,8-dihydrofolate | pH 7, 20°C, mutant enzyme V11D | Thermotoga maritima | |
| 1.5.1.3 | 0.05 | - |
NADPH | pH 7, 20°C, dimeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli prepared by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface | Escherichia coli | |
| 1.5.1.3 | 0.05 | - |
7,8-dihydrofolate | pH 7, 20°C, dimeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli prepared by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface | Escherichia coli | |
| 1.5.1.3 | 0.06 | - |
7,8-dihydrofolate | pH 7, 20°C, wild-type enzyme | Thermotoga maritima | |
| 1.5.1.3 | 1.39 | - |
NADPH | pH 7, 20°C, monomeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli prepared by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface | Escherichia coli | |
| 1.5.1.3 | 1.39 | - |
7,8-dihydrofolate | pH 7, 20°C, monomeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli prepared by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface | Escherichia coli | |
| 1.5.1.3 | 12.6 | - |
NADPH | pH 7, 20°C, wild-type enzyme | Escherichia coli | |
| 1.5.1.3 | 12.6 | - |
7,8-dihydrofolate | pH 7, 20°C, wild-type enzyme | Escherichia coli | |
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