Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 1.5.1.3 extracted from

  • Guo, J.; Loveridge, E.J.; Luk, L.Y.; Allemann, R.K.
    Effect of dimerization on dihydrofolate reductase catalysis (2013), Biochemistry, 52, 3881-3887 .
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
a variant (Xet-3) of dihydrofolate reductase from Escherichia coli constructed by introducing residues located at the Thermotoga maritima dihydrofolate reductase (TmDHFR) dimer interface is expressed in Escherichia coli BL21(DE3) cells Escherichia coli

Protein Variants

Protein Variants Comment Organism
additional information preparation of a dimeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli by introducing residues located at the Thermotoga maritima dihydrofolate reductase (TmDHFR) dimer interface increases the melting temperature to about 60°C, approximately 9°C higher than that measured for the Escherichia coli enzyme. The steady-state and pre-steady-state rate constants measured for Xet-3 were similar to those of dimeric TmDHFR but significantly lower than those of the parent dihydrofolate reductase from Escherichia coli Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0007
-
7,8-dihydrofolate pH 7, 20°C, wild-type enzyme Escherichia coli
0.0027
-
7,8-dihydrofolate pH 7, 20°C, monomeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli prepared by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface Escherichia coli
0.0048
-
NADPH pH 7, 20°C, wild-type enzyme Escherichia coli
0.0077
-
7,8-dihydrofolate pH 7, 20°C, dimeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli prepared by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface Escherichia coli
0.0195
-
NADPH pH 7, 20°C, monomeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli prepared by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface Escherichia coli
0.0199
-
NADPH pH 7, 20°C, dimeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli prepared by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface Escherichia coli

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
-
-
Thermotoga maritima Q60034
-
-
Thermotoga maritima DSM 3109 Q60034
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
7,8-dihydrofolate + NADPH + H+
-
Escherichia coli 5,6,7,8-tetrahydrofolate + NADP+
-
?
7,8-dihydrofolate + NADPH + H+
-
Thermotoga maritima 5,6,7,8-tetrahydrofolate + NADP+
-
?
7,8-dihydrofolate + NADPH + H+
-
Thermotoga maritima DSM 3109 5,6,7,8-tetrahydrofolate + NADP+
-
?

Subunits

Subunits Comment Organism
homodimer
-
Thermotoga maritima
monomer preparation of a dimeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface Escherichia coli

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
60
-
preparation of a dimeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface increases the melting temperature to about 60°C, approximately 9°C higher than that measured for the Escherichia coli enzyme Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.003
-
7,8-dihydrofolate pH 7, 20°C, mutant enzyme V11D Thermotoga maritima
0.05
-
NADPH pH 7, 20°C, dimeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli prepared by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface Escherichia coli
0.05
-
7,8-dihydrofolate pH 7, 20°C, dimeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli prepared by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface Escherichia coli
0.06
-
7,8-dihydrofolate pH 7, 20°C, wild-type enzyme Thermotoga maritima
1.39
-
NADPH pH 7, 20°C, monomeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli prepared by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface Escherichia coli
1.39
-
7,8-dihydrofolate pH 7, 20°C, monomeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli prepared by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface Escherichia coli
12.6
-
NADPH pH 7, 20°C, wild-type enzyme Escherichia coli
12.6
-
7,8-dihydrofolate pH 7, 20°C, wild-type enzyme Escherichia coli