Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli | Plasmodium falciparum |
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 | 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 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 | UniProt | Comment | Textmining |
---|---|---|---|
Plasmodium falciparum | - |
several Ser108 mutants | - |
Purification (Comment) | Organism |
---|---|
refolding and purification of wild-type and mutant enzymes from inclusion bodies in Escherichia coli | Plasmodium falciparum |
Renatured (Comment) | Organism |
---|---|
refolding of recombinant enzymes from inclusion bodies due to expression in E. coli | Plasmodium falciparum |
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 | 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 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 | Comment | Organism | Structure |
---|---|---|---|
NADPH | - |
Plasmodium falciparum |
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 |