Cloned (Comment) | Organism |
---|---|
the wild type and R653Q mutant pBKeDCS constructs are transformed into Escherichia coli BL21 DE3 to express the full-length protein | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
R653Q | mutant enzyme has normal substrate affinity but a 36% reduction in half-life at 42°C, the mutation reduces the metabolic activity of MTHFD1 within cells: formate incorporation into DNA in murine Mthfd1 knockout cells transfected with Arg653Gln is reduced by 26%, compared to cells transfected with wild-type protein, the mutant may increase the risk for neural tube defects, under ideal storage and standard assay conditions, the synthetase activity of MTHFD1 is unaffected by the R653Q mutation | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
ADP | - |
Homo sapiens | |
pteroylpentaglutamate | - |
Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0286 | - |
ATP | mutant R653Q | Homo sapiens | |
0.0302 | - |
ATP | wild type | Homo sapiens | |
0.0367 | - |
formate | wild type | Homo sapiens | |
0.0371 | - |
formate | mutant R653Q | Homo sapiens | |
0.333 | - |
tetrahydrofolate | mutant R653Q | Homo sapiens | |
0.364 | - |
tetrahydrofolate | wild type | Homo sapiens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
formate + ATP + tetrahydrofolate | Homo sapiens | synthetase activity of MTHFD1 | 10-formyltetrahydrofolate + ADP + phosphate | - |
? | |
additional information | Homo sapiens | (MTHFD1) is a trifunctional enzyme that interconverts tetrahydrofolate derivatives for nucleotide synthesis | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | P11586 | - |
- |
Purification (Comment) | Organism |
---|---|
by anion-exchange chromatography | Homo sapiens |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
100.2% activity of mutant R653Q after heating at 42°C for 90 min, in 7.5% glycerol and 0.35 mM folate pentaglutamate | Homo sapiens |
additional information | - |
102% activity of wild type enzyme after heating at 42°C for 90 min, in 7.5% glycerol and 0.35 mM folate pentaglutamate | Homo sapiens |
additional information | - |
57.4% activity of mutant R653Q, without adding of any cofactor, after heating at 42°C for 90 min, in 7.5% glycerol | Homo sapiens |
additional information | - |
72.3% activity of wild type enzyme, without adding of any cofactor, after heating at 42°C for 90 min, in 7.5% glycerol | Homo sapiens |
additional information | - |
80.5% activity of mutant R653Q after heating at 42°C for 90 min, in 7.5% glycerol, 1.2 mM MgCl, 0.6 mM ATP | Homo sapiens |
additional information | - |
88.2% activity of wild type enzyme after heating at 42°C for 90 min, in 7.5% glycerol, 1.2 mM MgCl, 0.6 mM ATP | Homo sapiens |
additional information | - |
enzyme knock-out cells are transfected with either wild type or R653Q mutant and their ability to synthesize purines is assessed by a 14C-formate incorporation assay, the cells expressing the R653Q variant incorporate only 74% of wild-type levels of formate into DNA, the R653Q protein appears to have a significant impact on cellular nucleotide metabolism | Homo sapiens |
23 | - |
synthetase activity of the wild type enzyme | Homo sapiens |
23.8 | - |
synthetase activity of the mutant R653Q | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(R,S)-tetrahydrofolate + ATP + formate | - |
Homo sapiens | ? | - |
r | |
formate + ATP + tetrahydrofolate | synthetase activity of MTHFD1 | Homo sapiens | 10-formyltetrahydrofolate + ADP + phosphate | - |
? | |
additional information | (MTHFD1) is a trifunctional enzyme that interconverts tetrahydrofolate derivatives for nucleotide synthesis | Homo sapiens | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
Formyltetrahydrofolate synthetase | - |
Homo sapiens |
methylenetetrahydrofolate dehydrogenase-methenyltetrahydrofolate cyclohydrolase-formyltetrahydrofolate synthetase | - |
Homo sapiens |
MTHFD1 | - |
Homo sapiens |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
additional information | - |
thermolability is reduced by magnesium adenosine triphosphate and eliminated by the substrate analog folate pentaglutamate, suggesting that folate status may modulate impact of the variant R653Q | Homo sapiens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Homo sapiens |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0296 | - |
pteroylpentaglutamate | wild type enzyme | Homo sapiens | |
0.0328 | - |
pteroylpentaglutamate | mutant R653Q | Homo sapiens | |
0.0338 | - |
ADP | mutant R653Q | Homo sapiens | |
0.0364 | - |
ADP | wild type enzyme | Homo sapiens |