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Literature summary extracted from

  • Delves, C.J.; Ballantine, S.P.; Tansik, R.L.; Baccanari, D.P.; Stammers, D.K.
    Refolding of recombinant Pneumocystis carinii dihydrofolate reductase and characterization of the enzyme (1993), Protein Expr. Purif., 4, 16-23.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.5.1.3 Cl-
-
Pneumocystis carinii
1.5.1.3 KCl
-
Rattus norvegicus
1.5.1.3 KCl recombinant enzyme, concentration 0.1 M Pneumocystis carinii

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.5.1.3 rat-derived gene is overexpressed in Escherichia coli Pneumocystis carinii

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.5.1.3 crystallization of refolded recombinant enzyme as ternary complex with NADPH and various inhibitors Pneumocystis carinii

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.5.1.3 KCl recombinant enzyme, concentration above 0.1 M Pneumocystis carinii
1.5.1.3 methotrexate recombinant enzyme Pneumocystis carinii
1.5.1.3 methotrexate
-
Rattus norvegicus
1.5.1.3 piritrexim recombinant enzyme Pneumocystis carinii
1.5.1.3 pyrimethamine
-
Pneumocystis carinii
1.5.1.3 trimethoprim
-
Pneumocystis carinii
1.5.1.3 trimetrexate recombinant enzyme Pneumocystis carinii
1.5.1.3 Urea recombinant enzyme, above 4 M Pneumocystis carinii

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.5.1.3 additional information
-
additional information buffer and salt conditions significantly alter the Km-values for the substrates Pneumocystis carinii
1.5.1.3 0.0023
-
7,8-dihydrofolate recombinant enzyme, pH 7.0, 0.1 M imidazole Pneumocystis carinii
1.5.1.3 0.003
-
NADPH recombinant enzyme, pH 7.0, 0.1 M imidazole Pneumocystis carinii

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
1.5.1.3 soluble recombinant enzyme expressed in Escherichia coli Pneumocystis carinii
-
-

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.5.1.3 24000
-
recombinant enzyme, gel filtration Pneumocystis carinii

Organism

EC Number Organism UniProt Comment Textmining
1.5.1.3 Pneumocystis carinii
-
rat-derived
-
1.5.1.3 Rattus norvegicus
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.5.1.3 recombinant purified from Escherichia coli Pneumocystis carinii

Renatured (Commentary)

EC Number Renatured (Comment) Organism
1.5.1.3 refolding after solubilisation from inclusion bodies with 4 M guanidium hydrochloride in 0.5% polyethylene glycol 1450, pH 7.0 Pneumocystis carinii

Source Tissue

EC Number Source Tissue Comment Organism Textmining
1.5.1.3 lung source of Pneumocystis carnii Rattus norvegicus
-
1.5.1.3 additional information derived from rat lung Pneumocystis carinii
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.5.1.3 42
-
recombinant enzyme, partially purified, after refolding Pneumocystis carinii
1.5.1.3 321
-
purified recombinant enzyme Pneumocystis carinii

Storage Stability

EC Number Storage Stability Organism
1.5.1.3 -20°C, 50% glycerol, 30% loss of activity after 9 months Pneumocystis carinii
1.5.1.3 -70°C, 10% glycerol, less than 5% loss of activity after 9 months Pneumocystis carinii

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.5.1.3 7,8-dihydrofolate + NADPH
-
Rattus norvegicus 5,6,7,8-tetrahydrofolate + NADP+
-
?
1.5.1.3 7,8-dihydrofolate + NADPH
-
Pneumocystis carinii 5,6,7,8-tetrahydrofolate + NADP+
-
?
1.5.1.3 folate + NADPH no activity Pneumocystis carinii 5,6,7,8-tetrahydrofolate + NADP+
-
?

Subunits

EC Number Subunits Comment Organism
1.5.1.3 monomer
-
Rattus norvegicus
1.5.1.3 monomer 1 * 24000, recombinant enzyme, SDS-PAGE Pneumocystis carinii

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.5.1.3 30
-
assay at Pneumocystis carinii

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.5.1.3 108
-
7,8-dihydrofolate recombinant enzyme Pneumocystis carinii

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.5.1.3 7.5
-
sharp optimum, recombinant enzyme Pneumocystis carinii

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.5.1.3 6 9 recombinant, refolded enzyme Pneumocystis carinii

Cofactor

EC Number Cofactor Comment Organism Structure
1.5.1.3 NADPH
-
Rattus norvegicus
1.5.1.3 NADPH
-
Pneumocystis carinii

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.5.1.3 0.00000000002
-
methotrexate recombinant enzyme Pneumocystis carinii
1.5.1.3 0.0007
-
pyrimethamine recombinant enzyme Pneumocystis carinii