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Literature summary for 6.3.4.3 extracted from

  • Christensen, K.E.; Rohlicek, C.V.; Andelfinger, G.U.; Michaud, J.; Bigras, J.L.; Richter, A.; Mackenzie, R.E.; Rozen, R.
    The MTHFD1 p.Arg653Gln variant alters enzyme function and increases risk for congenital heart defects (2009), Hum. Mutat., 30, 212-220.
    View publication on PubMed
Search for more literature for 6.3.4.3

Cloned(Commentary)

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

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

Inhibitors Comment Organism Structure
ADP
-
 Homo sapiens
pteroylpentaglutamate
-
 Homo sapiens

KM Value [mM]

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/ Products (Substrates)

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

Organism UniProt Comment Textmining
Homo sapiens P11586
-
-

Purification (Commentary)

Purification (Comment) Organism
by anion-exchange chromatography Homo sapiens

Specific Activity [micromol/min/mg]

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 and Products (Substrate)

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

Synonyms Comment Organism
Formyltetrahydrofolate synthetase
-
 Homo sapiens
methylenetetrahydrofolate dehydrogenase-methenyltetrahydrofolate cyclohydrolase-formyltetrahydrofolate synthetase
-
 Homo sapiens
MTHFD1
-
 Homo sapiens

Temperature Stability [°C]

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

Cofactor Comment Organism Structure
ATP
-
 Homo sapiens

Ki Value [mM]

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