Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Thermus thermophilus |
Crystallization (Comment) | Organism |
---|---|
hanging drop vapor diffusion method, using 0.1 M sodium acetate buffer (NaOAc, pH 4.3,4.5), 1 M lithium chloride, 10% (w/v) polyethylene glycol 6000, 10-20% (v/v) glycerol, and 2-5% (v/v) dioxane, at 20°C | Thermus thermophilus |
Protein Variants | Comment | Organism |
---|---|---|
A222V | the mutant shows enhanced instability upon loss of FAD | Thermus thermophilus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.18 | - |
(6R)-5,10-methylenetetrahydrofolate | at pH 7.0 and 50°C | Thermus thermophilus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(6R)-5,10-methylenetetrahydrofolate + NAD(P)H + H+ | Thermus thermophilus | - |
(6S)-5-methyltetrahydrofolate + NAD(P)+ | - |
r | |
(6R)-5,10-methylenetetrahydrofolate + NAD(P)H + H+ | Thermus thermophilus HB8 / ATCC 27634 / DSM 579 | - |
(6S)-5-methyltetrahydrofolate + NAD(P)+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermus thermophilus | Q5SLG6 | - |
- |
Thermus thermophilus HB8 / ATCC 27634 / DSM 579 | Q5SLG6 | - |
- |
Purification (Comment) | Organism |
---|---|
Toyopearl SuperQ-650M column chromatography and nickel affinity column chromatography | Thermus thermophilus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(6R)-5,10-methylenetetrahydrofolate + NAD(P)H + H+ | - |
Thermus thermophilus | (6S)-5-methyltetrahydrofolate + NAD(P)+ | - |
r | |
(6R)-5,10-methylenetetrahydrofolate + NAD(P)H + H+ | - |
Thermus thermophilus HB8 / ATCC 27634 / DSM 579 | (6S)-5-methyltetrahydrofolate + NAD(P)+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
heterodimer | x-ray crystallography | Thermus thermophilus |
Synonyms | Comment | Organism |
---|---|---|
MTHFR | - |
Thermus thermophilus |
NADH:CH2-H4folate oxidoreductase | - |
Thermus thermophilus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | the enzyme contains one FAD prosthetic group bound per dimer. Km for FAD is 0.005 mM. The enzyme activity of the FAD-replete enzyme is approximately 50% compared to the normal purified enzyme | Thermus thermophilus | |
NADH | - |
Thermus thermophilus | |
NADPH | - |
Thermus thermophilus |