Literature summary for 1.5.1.3 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.

Search for more literature for 1.5.1.3

Application

Application	Comment	Organism
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

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

Inhibitors

Inhibitors	Comment	Organism	Structure
cycloguanil	-	Plasmodium falciparum
pyrimethamine	-	Plasmodium falciparum

KM Value [mM]

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

Organism

Organism	UniProt	Comment	Textmining
Plasmodium falciparum	P13922	-	-

Purification (Commentary)

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

Substrates and Products (Substrate)

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

Ki Value [mM]

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

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Release 2023.1 (January 2023)