Literature summary for 1.5.1.3 extracted from

Sirawaraporn, W.; Yongkiettrakul, S.; Sirawaraporn, R.; Yuthavong, Y.; Santi, D.V.
Plasmodium falciparum: asparagine mutant at residue 108 of dihydrofolate reductase is an optimal antifolate-resistant single mutant (1997), Exp. Parasitol., 87, 245-252.

Search for more literature for 1.5.1.3

Cloned(Commentary)

Cloned (Comment)	Organism
expression of wild-type and mutant enzymes in Escherichia coli	Plasmodium falciparum

Protein Variants

Protein Variants	Comment	Organism
S108A	combinatorial mutagenesis	Plasmodium falciparum
S108C	combinatorial mutagenesis	Plasmodium falciparum
S108D	combinatorial mutagenesis, no detectable activity on refolding from inclusion bodies	Plasmodium falciparum
S108E	combinatorial mutagenesis, no detectable activity on refolding from inclusion bodies	Plasmodium falciparum
S108F	combinatorial mutagenesis, no detectable activity on refolding from inclusion bodies	Plasmodium falciparum
S108G	combinatorial mutagenesis	Plasmodium falciparum
S108H	combinatorial mutagenesis, no detectable activity on refolding from inclusion bodies	Plasmodium falciparum
S108I	combinatorial mutagenesis, no detectable activity on refolding from inclusion bodies	Plasmodium falciparum
S108L	combinatorial mutagenesis, reduced activity	Plasmodium falciparum
S108M	combinatorial mutagenesis, reduced activity	Plasmodium falciparum
S108N	combinatorial mutagenesis, conferred resistance to pyrimethamine and cycloguanil	Plasmodium falciparum
S108P	combinatorial mutagenesis, no detectable activity on refolding from inclusion bodies	Plasmodium falciparum
S108Q	combinatorial mutagenesis, reduced activity, conferred resistance to pyrimethamine and cycloguanil	Plasmodium falciparum
S108R	combinatorial mutagenesis, no detectable activity on refolding from inclusion bodies	Plasmodium falciparum
S108T	combinatorial mutagenesis, reduced activity	Plasmodium falciparum
S108V	combinatorial mutagenesis, reduced activity	Plasmodium falciparum
S108W	combinatorial mutagenesis, no detectable activity on refolding from inclusion bodies	Plasmodium falciparum
S108Y	combinatorial mutagenesis, no detectable activity on refolding from inclusion bodies	Plasmodium falciparum

Inhibitors

Inhibitors	Comment	Organism	Structure
cycloguanil	wild-type and several Ser108 mutant enzymes	Plasmodium falciparum
pyrimethamine	wild-type and several mutants of Ser108	Plasmodium falciparum

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.005	-	NADPH	recombinant wild-type enzyme	Plasmodium falciparum
0.007	-	NADPH	recombinant mutant S108N	Plasmodium falciparum
0.009	-	NADPH	recombinant mutants S108G, S108A and S108C	Plasmodium falciparum
0.013	-	7,8-dihydrofolate	recombinant wild-type enzyme	Plasmodium falciparum
0.014	-	7,8-dihydrofolate	recombinant mutants S108T and S108A	Plasmodium falciparum
0.014	-	NADPH	recombinant mutant S108Q	Plasmodium falciparum
0.017	-	NADPH	recombinant mutant S108T	Plasmodium falciparum
0.02	0.025	7,8-dihydrofolate	recombinant mutants S108G, S108N and S108C	Plasmodium falciparum
0.05	-	7,8-dihydrofolate	recombinant mutant S108Q	Plasmodium falciparum

Organism

Organism	UniProt	Comment	Textmining
Plasmodium falciparum	-	several Ser108 mutants	-

Purification (Commentary)

Purification (Comment)	Organism
refolding and purification of wild-type and mutant enzymes from inclusion bodies in Escherichia coli	Plasmodium falciparum

Renatured (Commentary)

Renatured (Comment)	Organism
refolding of recombinant enzymes from inclusion bodies due to expression in E. coli	Plasmodium falciparum

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
0.6	0.7	recombinant mutants S108V and S108M	Plasmodium falciparum
1	-	recombinant mutants S108C and S108L	Plasmodium falciparum
2	-	recombinant mutant S108A	Plasmodium falciparum
4	-	recombinant mutant S108G	Plasmodium falciparum
5	-	recombinant mutant S108Q	Plasmodium falciparum
8	-	recombinant mutant S108T	Plasmodium falciparum
9	-	recombinant mutant S108N	Plasmodium falciparum
17	-	recombinant wild-type enzyme	Plasmodium falciparum

Storage Stability

Storage Stability	Organism
-20°C, 20 mM potassium phosphate, pH 7.0, 0.1 mM EDTA, 10 mM dithiothreitol, 0.2 M KCl, 20% glycerol, mutant S108Q loss of 40% activity after 7 months	Plasmodium falciparum
-20°C, 20 mM potassium phosphate, pH7.0, 0.1 mM EDTA, 10 mM dithiothreitol, 0.2 M KCl, 20% glycerol, mutants S108G, S108A and S108C stable over 7 months	Plasmodium falciparum

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
12	-	7,8-dihydrofolate	recombinant mutant S108C	Plasmodium falciparum
21	-	7,8-dihydrofolate	recombinant mutant S108A	Plasmodium falciparum
23	-	7,8-dihydrofolate	recombinant mutant S108G	Plasmodium falciparum
36	-	7,8-dihydrofolate	recombinant mutant S108T	Plasmodium falciparum
37	-	7,8-dihydrofolate	recombinant mutant S108Q	Plasmodium falciparum
88	-	7,8-dihydrofolate	recombinant wild-type enzyme	Plasmodium falciparum
92	-	7,8-dihydrofolate	recombinant mutant S108N	Plasmodium falciparum

Cofactor

Cofactor	Comment	Organism	Structure
NADPH	-	Plasmodium falciparum

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism	Structure
0.0000014	-	pyrimethamine	recombinant mutant S108T	Plasmodium falciparum
0.0000015	-	pyrimethamine	recombinant wild-type enzyme	Plasmodium falciparum
0.0000016	-	cycloguanil	recombinant mutant S108T	Plasmodium falciparum
0.000002	-	cycloguanil	recombinant mutant S108A	Plasmodium falciparum
0.0000026	-	cycloguanil	recombinant wild-type enzyme	Plasmodium falciparum
0.000003	-	cycloguanil	recombinant mutant S108C	Plasmodium falciparum
0.000004	-	pyrimethamine	recombinant mutant S108A	Plasmodium falciparum
0.000006	-	pyrimethamine	recombinant mutant S108C	Plasmodium falciparum
0.000007	-	pyrimethamine	recombinant mutant S108G	Plasmodium falciparum
0.000013	-	pyrimethamine	recombinant mutant S108N	Plasmodium falciparum
0.000015	-	cycloguanil	recombinant mutant S108N	Plasmodium falciparum
0.000017	-	cycloguanil	recombinant mutant S108G	Plasmodium falciparum
0.000095	-	pyrimethamine	recombinant mutant S108Q	Plasmodium falciparum
0.000158	-	cycloguanil	recombinant mutant S108Q	Plasmodium falciparum

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Release 2023.1 (January 2023)