Cloned (Comment) | Organism |
---|---|
expression in Sf9 insect cells | Homo sapiens |
General Stability | Organism |
---|---|
the enzyme precipitates after a week at concentrations as low as 0.1 mg/ml | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00123 | - |
5,6,7,8-tetrahydrofolate-Glu2 | pH 8.7 | Homo sapiens | |
0.0104 | - |
aminopterin | pH 8.7 | Homo sapiens | |
0.04 | - |
methotrexate | pH 8.7 | Homo sapiens | |
0.046 | - |
ATP | pH 8.7 | Homo sapiens | |
0.126 | - |
N-(p(((2-amino-4-hydroxy-6-pteridinyl)methyl)-amino)benzoyl)glutamic acid | pH 8.7 | Homo sapiens | |
0.24 | - |
L-glutamate | pH 8.7 | Homo sapiens |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
cytosol | - |
Homo sapiens | 5829 | - |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Homo sapiens | the enzyme catalyzes the formation of an amide bond between glutamic acid and the gamma-carboxyl group of any of the naturally occurring folate compounds | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | - |
human | - |
Purification (Comment) | Organism |
---|---|
ammonium sulfate fractionation, chromatography on Sephacryl S-100HR and DEAE-Sephacel ion exchange column, 25fold purification | Homo sapiens |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
CEM cell | leukemic cells, cDNA, recombinant enzyme | Homo sapiens | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.0083 | - |
supernatant of cell extract | Homo sapiens |
0.016 | - |
ammonium sulfate fraction | Homo sapiens |
0.13 | - |
Sephacryl S-100 HR fraction | Homo sapiens |
0.22 | - |
DEAE-Sephacel fraction | Homo sapiens |
Storage Stability | Organism |
---|---|
-20ºC, 0.05% n-octylglucoside, DTT 2 mM, 50% glycerol, stable for 9 months. Replenishment of DTT every 2 months is necesary. | Homo sapiens |
0ºC, leupeptin, MgATP2- 2 mM and 2-mercaptoethanol 50 mM or DTT 2 mM, limited stability. More stable with DTT in place of 2-mercaptoethanol. | Homo sapiens |
20ºC, Tris buffer, without thiols, 85% activity over 30 min. | Homo sapiens |
37ºC, Tris buffer, without thiols, with or without MgATP2- 5 mM, highly unstable, half-life of 6-7.3 min. Addition of 2-mercaptoethanol increases half-life of the enzyme to 11-21 min. Addition of 0.3 mg/ml BSA, stabilizes for 2 hours. | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + 5,10-dideazatetrahydrofolic acid + L-glutamate | - |
Homo sapiens | ADP + 5,10-dideazatetrahydrofolyl-L-Glu + phosphate | - |
? | |
ATP + 5,6,7,8-tetrahydrofolate-Glu2 + L-glutamate | - |
Homo sapiens | ADP + phosphate + 5,6,7,8-tetrahydrofolyl-Glu3 | - |
? | |
ATP + aminopterin + L-glutamate | - |
Homo sapiens | ? | - |
? | |
ATP + methotrexate + Glu | - |
Homo sapiens | ADP + phosphate + methotrexyl-Glu | - |
? | |
ATP + methotrexate-Glu + L-glutamate | - |
Homo sapiens | ADP + methotrexate-Glu2 + phosphate | - |
? | |
ATP + N-(p(((2-amino-4-hydroxy-6-pteridinyl)methyl)-amino)benzoyl)glutamic acid + L-glutamate | folic acid, poor substrate | Homo sapiens | ADP + phosphate + N-(p(((2-amino-4-hydroxy-6-pteridinyl)methyl)-amino)benzoyl)Glu2 | - |
? | |
additional information | the enzyme catalyzes the formation of an amide bond between glutamic acid and the gamma-carboxyl group of any of the naturally occurring folate compounds | Homo sapiens | ? | - |
? |