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Literature summary for 1.8.1.4 extracted from

  • Laine, L.M.; Biddau, M.; Byron, O.; Mueller, S.
    Biochemical and structural characterization of the apicoplast dihydrolipoamide dehydrogenase of Plasmodium falciparum (2015), Biosci. Rep., 35, e00171.
    View publication on PubMedView publication on EuropePMC

Application

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(Commentary)

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

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 [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

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 [Da] Molecular Weight Maximum [Da] Comment Organism
127478
-
sequence calculation Plasmodium falciparum
128000 140000 gel filtration and analytical ultracentrifugation Plasmodium falciparum

Natural Substrates/ Products (Substrates)

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

Organism UniProt Comment Textmining
Plasmodium falciparum
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme from Escherichia coli strain NovaBlue (DE3) by nickel affinity chromatography and gel filtration Plasmodium falciparum

Substrates and Products (Substrate)

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

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

Synonyms Comment Organism
apicoplast E3
-
Plasmodium falciparum
dihydrolipoamide dehydrogenase
-
Plasmodium falciparum
E3
-
Plasmodium falciparum
PfaE3
-
Plasmodium falciparum

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Plasmodium falciparum

Turnover Number [1/s]

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

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

Cofactor Comment Organism Structure
NAD+
-
Plasmodium falciparum
NADH
-
Plasmodium falciparum

General Information

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