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Literature summary for 6.3.2.17 extracted from

  • Sanghani, P.C.; Moran, R.G.
    Purification and characteristics of recombinant human folylpoly-gamma-glutamate synthetase expressed at high levels in insect cells (2000), Protein Expr. Purif., 18, 36-45.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression in Sf9 insect cells Homo sapiens

General Stability

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 [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

Localization Comment Organism GeneOntology No. Textmining
cytosol
-
Homo sapiens 5829
-

Natural Substrates/ Products (Substrates)

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

Organism UniProt Comment Textmining
Homo sapiens
-
human
-

Purification (Commentary)

Purification (Comment) Organism
ammonium sulfate fractionation, chromatography on Sephacryl S-100HR and DEAE-Sephacel ion exchange column, 25fold purification Homo sapiens

Source Tissue

Source Tissue Comment Organism Textmining
CEM cell leukemic cells, cDNA, recombinant enzyme Homo sapiens
-

Specific Activity [micromol/min/mg]

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

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 and Products (Substrate)

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 ?
-
?