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Literature summary extracted from

  • Japrung, D.; Chusacultanachai, S.; Yuvaniyama, J.; Wilairat, P.; Yuthavong, Y.
    A simple dual selection for functionally active mutants of Plasmodium falciparum dihydrofolate reductase with improved solubility (2005), Protein Eng. Des. Sel., 18, 457-464.
    View publication on PubMed

Application

EC Number Application Comment Organism
1.5.1.3 biotechnology in vivo screening system to select for functionally active proteins with increased solubility. Fusion of enzyme to green fluorescent protein as reporter for solubility Plasmodium falciparum

Protein Variants

EC Number Protein Variants Comment Organism
1.5.1.3 K27E kinetic properties similar to wild-type, increase in solubility Plasmodium falciparum
1.5.1.3 Y35G/F37L kinetic properties similar to wild-type, increase in solubility Plasmodium falciparum
1.5.1.3 Y35L/F37R kinetic properties similar to wild-type, increase in solubility Plasmodium falciparum
1.5.1.3 Y35L/F37T kinetic properties similar to wild-type, increase in solubility Plasmodium falciparum
1.5.1.3 Y35Q/F37R kinetic properties similar to wild-type, increase in solubility Plasmodium falciparum

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.5.1.3 cycloguanil
-
 Plasmodium falciparum
1.5.1.3 pyrimethamine
-
 Plasmodium falciparum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.5.1.3 0.0042
-
 7,8-dihydrofolate wild-type, pH 5.0, 25°C Plasmodium falciparum
1.5.1.3 0.0043
-
 NADPH mutant Y35G/F37L, pH 5.0, 25°C Plasmodium falciparum
1.5.1.3 0.0048
-
 NADPH mutant Y35L/F37T, pH 5.0, 25°C Plasmodium falciparum
1.5.1.3 0.0049
-
 7,8-dihydrofolate mutant Y35G/F37L, pH 5.0, 25°C Plasmodium falciparum
1.5.1.3 0.0053
-
 7,8-dihydrofolate mutant K27E, pH 5.0, 25°C Plasmodium falciparum
1.5.1.3 0.0055
-
 7,8-dihydrofolate mutant Y35L/F37R, pH 5.0, 25°C Plasmodium falciparum
1.5.1.3 0.0058
-
 NADPH wild-type, pH 5.0, 25°C Plasmodium falciparum
1.5.1.3 0.0068
-
 NADPH mutant K27E, pH 5.0, 25°C Plasmodium falciparum
1.5.1.3 0.0076
-
 7,8-dihydrofolate mutant Y35Q/F37R, pH 5.0, 25°C Plasmodium falciparum
1.5.1.3 0.0086
-
 7,8-dihydrofolate mutant Y35L/F37T, pH 5.0, 25°C Plasmodium falciparum
1.5.1.3 0.0097
-
 NADPH mutant Y35L/F37R, pH 5.0, 25°C Plasmodium falciparum
1.5.1.3 0.0105
-
 NADPH mutant Y35Q/F37R, pH 5.0, 25°C Plasmodium falciparum

Organism

EC Number Organism UniProt Comment Textmining
1.5.1.3 Plasmodium falciparum P13922
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.5.1.3 wild-type enzyme readily aggregates at 2 mg/ml Plasmodium falciparum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.5.1.3 7,8-dihydrofolate + NADPH
-
 Plasmodium falciparum 5,6,7,8-tetrahydrofolate + NADP+
-
 ?

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.5.1.3 0.00000012
-
 pyrimethamine mutant K27E, pH 5.0, 25°C Plasmodium falciparum
1.5.1.3 0.0000002
-
 pyrimethamine mutant Y35G/F37L, pH 5.0, 25°C Plasmodium falciparum
1.5.1.3 0.00000024
-
 cycloguanil mutant K27E, pH 5.0, 25°C Plasmodium falciparum
1.5.1.3 0.00000028
-
 cycloguanil mutant Y35G/F37L, pH 5.0, 25°C Plasmodium falciparum
1.5.1.3 0.00000057
-
 pyrimethamine mutant Y35L/F37T, pH 5.0, 25°C Plasmodium falciparum
1.5.1.3 0.0000006
-
 cycloguanil mutant Y35Q/F37R, pH 5.0, 25°C Plasmodium falciparum
1.5.1.3 0.0000006
-
 pyrimethamine wild-type, pH 5.0, 25°C Plasmodium falciparum
1.5.1.3 0.00000061
-
 pyrimethamine mutant Y35Q/F37R, pH 5.0, 25°C Plasmodium falciparum
1.5.1.3 0.0000012
-
 pyrimethamine mutant Y35L/F37R, pH 5.0, 25°C Plasmodium falciparum
1.5.1.3 0.0000012
-
 cycloguanil mutant Y35L/F37R, pH 5.0, 25°C Plasmodium falciparum
1.5.1.3 0.0000015
-
 cycloguanil wild-type, pH 5.0, 25°C Plasmodium falciparum
1.5.1.3 0.0000018
-
 cycloguanil mutant Y35L/F37T, pH 5.0, 25°C Plasmodium falciparum