Application | Comment | Organism |
---|---|---|
drug development | a homology model for PfaE3 reveals an extra anti-parallel beta-strand at the position where human E3BP (E3-binding protein) interacts with E3, a parasite-specific feature that may be exploitable for drug discovery against pyruvate dehydrogenase complex, PDC. Plasmodium PDC is essential for parasite survival in the mosquito vector and for late liver stage development in the human host, suggesting its suitability as a target for intervention strategies against malaria | Plasmodium falciparum |
Cloned (Comment) | Organism |
---|---|
gene Pfae3, recombinant expression of His-tagged enzyme in Escherichia coli strain NovaBlue (DE3), transfection of pCC1-Pfae3 Plasmodium falciparum 3D7 erythrocytic stages | Plasmodium falciparum |
Protein Variants | Comment | Organism |
---|---|---|
additional information | generation of Pfae3 deletion mutants of Plasmodium falciparum, the gene is replaced by the selectable marker hdhfr after initial positive selection with WR99210 followed by negative selection using 5-fluorocytosine generating the line 3D7DELTAae3, phenotype, overview | Plasmodium falciparum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.015 | - |
NADH | pH 7.0, 25°C, recombinant His-tagged enzyme | Plasmodium falciparum | |
0.096 | - |
NAD+ | pH 8.0, 25°C, recombinant His-tagged enzyme | Plasmodium falciparum | |
0.841 | - |
Lipoamide | pH 7.0, 25°C, recombinant His-tagged enzyme | Plasmodium falciparum | |
1.16 | - |
dihydrolipoamide | pH 8.0, 25°C, recombinant His-tagged enzyme | Plasmodium falciparum |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
apicoplast | in the human malaria parasite Plasmodium falciparum, the single PDC enzyme complex with enzyme E3 is located exclusively in the apicoplast | Plasmodium falciparum | 20011 | - |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
127478 | - |
sequence calculation | Plasmodium falciparum |
128000 | 140000 | gel filtration and analytical ultracentrifugation | Plasmodium falciparum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
dihydrolipoamide + NAD+ | Plasmodium falciparum | - |
lipoamide + NADH + H+ | - |
r | |
lipoamide + NADH + H+ | Plasmodium falciparum | - |
dihydrolipoamide + NAD+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Plasmodium falciparum | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli strain NovaBlue (DE3) by nickel affinity chromatography and gel filtration | Plasmodium falciparum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
dihydrolipoamide + NAD+ | - |
Plasmodium falciparum | lipoamide + NADH + H+ | - |
r | |
lipoamide + NADH + H+ | - |
Plasmodium falciparum | dihydrolipoamide + NAD+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 64000, recombinant Hs-tagged enzyme, SDS-PAGE | Plasmodium falciparum |
More | E3 enzyme homology structure modelling using the human enzyme structure, PDB ID 2F5Z | Plasmodium falciparum |
Synonyms | Comment | Organism |
---|---|---|
apicoplast E3 | - |
Plasmodium falciparum |
dihydrolipoamide dehydrogenase | - |
Plasmodium falciparum |
E3 | - |
Plasmodium falciparum |
PfaE3 | - |
Plasmodium falciparum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Plasmodium falciparum |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
77.8 | - |
NADH | pH 7.0, 25°C, recombinant His-tagged enzyme | Plasmodium falciparum | |
97 | - |
Lipoamide | pH 7.0, 25°C, recombinant His-tagged enzyme | Plasmodium falciparum | |
110 | - |
NAD+ | pH 8.0, 25°C, recombinant His-tagged enzyme | Plasmodium falciparum | |
110 | - |
dihydrolipoamide | pH 8.0, 25°C, recombinant His-tagged enzyme | Plasmodium falciparum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at, reduction of lipoamide with NADH | Plasmodium falciparum |
8 | - |
assay at, oxidation of dihydrolipoamide with NAD+ | Plasmodium falciparum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Plasmodium falciparum | |
NADH | - |
Plasmodium falciparum |
General Information | Comment | Organism |
---|---|---|
malfunction | Pfae3 is deleted from Plasmodium falciparum and although the mutants are viable, they display a highly synchronous growth phenotype during intra-erythrocytic development. The mutants also show changes in the expression of some mitochondrial and antioxidant proteins suggesting that deletion of Pfae3 impacts on the parasite's metabolic function with downstream effects on the parasite's redox homoeostasis and cell cycle | Plasmodium falciparum |
additional information | a homology model for PfaE3 reveals an extra anti-parallel beta-strand at the position where human E3BP (E3-binding protein) interacts with E3, a parasite-specific feature that may be exploitable for drug discovery against PDC. E3 enzyme homology structure modelling using the human enzyme structure, PDB ID 2F5Z | Plasmodium falciparum |
physiological function | pyruvate dehydrogenase complex, PDC, is a multi-enzyme complex comprising an E1, pyruvate decarboxylase, an E2, dihydrolipomide acetyltransferase, and an E3, dihydrolipoamide dehydrogenase. Plasmodium PDC is essential for parasite survival in the mosquito vector and for late liver stage development in the human host | Plasmodium falciparum |