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Literature summary for 1.8.1.4 extracted from

  • McMillan, P.J.; Stimmler, L.M.; Foth, B.J.; McFadden, G.I.; Mueller, S.
    The human malaria parasite Plasmodium falciparum possesses two distinct dihydrolipoamide dehydrogenases (2005), Mol. Microbiol., 55, 27-38.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
DNA and amino acid sequence determination and analysis of both lipdh genes using RT-PCR, expression of the mitochondrial isozyme as GFP-fusion protein giving green fluorescence and of the apicoplast isozyme fused to acyl-carrier-protein resulting in red fluorescence, expression of truncated mitochondrial and apicoplastic isozymes lacking the putative target sequences as His-tagged proteins in Escherichia coli strain BL21-RIL(DE3), the apicoplast isozyme is expressed at very low levels Plasmodium falciparum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information ping pong kinetics Plasmodium falciparum
0.021
-
NADH pH 8.0, 25°C, mitochondrial isozyme Plasmodium falciparum
0.045
-
NAD+ pH 8.0, 25°C, mitochondrial isozyme Plasmodium falciparum
0.146
-
dihydrolipoamide pH 8.0, 25°C, mitochondrial isozyme Plasmodium falciparum
0.87
-
Lipoamide pH 8.0, 25°C, mitochondrial isozyme Plasmodium falciparum

Localization

Localization Comment Organism GeneOntology No. Textmining
apicoplast isozyme aLipDH Plasmodium falciparum 20011
-
mitochondrion isozyme mLipDH is a component of the branched-chain 2-ketoacid dehydrogenase and the 2-ketoglutarate dehydrogenase multienzyme complexes Plasmodium falciparum 5739
-
additional information subcellular localization study using recombinant expression of the isozymes fused to fluorescent protein tags Plasmodium falciparum
-
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
57200
-
2 * 57200, mitochondrial enzyme, SDS-PAGE and gel filtration, 2 * 75600, apicoplast enzyme, SDS-PAGE and gel filtration Plasmodium falciparum
75600
-
2 * 57200, mitochondrial enzyme, SDS-PAGE and gel filtration, 2 * 75600, apicoplast enzyme, SDS-PAGE and gel filtration Plasmodium falciparum

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
dihydrolipoamide + NAD+ Plasmodium falciparum the forward reaction is the physiological one lipoamide + NADH
-
r

Organism

Organism UniProt Comment Textmining
Plasmodium falciparum
-
3D7, 2 lipdh genes encoding 2 isozymes in mitochondrion and apicoplast, both indispensable components of the 2-ketoacid dehydrogenase multienzyme complexes
-

Posttranslational Modification

Posttranslational Modification Comment Organism
additional information the apicoplast isozyme contains a potential transit peptide, the mitochondrial isozyme contains targeting sequence Plasmodium falciparum

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged truncated isozymes from Escherichia coli by nickel affinity chromatography to 98% homogeneity Plasmodium falciparum

Reaction

Reaction Comment Organism Reaction ID
protein N6-(dihydrolipoyl)lysine + NAD+ = protein N6-(lipoyl)lysine + NADH + H+ the mitochondrial isozyme shows ping pong kinetic mechanism Plasmodium falciparum

Source Tissue

Source Tissue Comment Organism Textmining
additional information both genes are transcribed during erythrocytic development of the parasite Plasmodium falciparum
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
dihydrolipoamide + NAD+
-
Plasmodium falciparum lipoamide + NADH
-
r
dihydrolipoamide + NAD+ the forward reaction is the physiological one Plasmodium falciparum lipoamide + NADH
-
r

Subunits

Subunits Comment Organism
dimer 2 * 57200, mitochondrial enzyme, SDS-PAGE and gel filtration, 2 * 75600, apicoplast enzyme, SDS-PAGE and gel filtration Plasmodium falciparum
More 2 distinct dihydrolipoamide dehydrogenases, both indispensable components of the 2-ketoacid dehydrogenase multienzyme complexes Plasmodium falciparum

Synonyms

Synonyms Comment Organism
dihydrolipoamide dehydrogenase
-
Plasmodium falciparum
LipDH
-
Plasmodium falciparum

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
135.5 337 NAD+ pH 8.0, 25°C, mitochondrial isozyme, forward reaction, varying conditions Plasmodium falciparum
135.5 337 Lipoamide pH 8.0, 25°C, mitochondrial isozyme, forward reaction, varying conditions Plasmodium falciparum
448
-
NADH pH 8.0, 25°C, mitochondrial isozyme, reverse reaction, varying conditions Plasmodium falciparum
448
-
dihydrolipoamide pH 8.0, 25°C, mitochondrial isozyme, reverse reaction, varying conditions Plasmodium falciparum

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
reverse reaction, isozyme mLipDH Plasmodium falciparum
9
-
forward reaction, isozyme LipDH Plasmodium falciparum

Cofactor

Cofactor Comment Organism Structure
FAD protein-bound Plasmodium falciparum
NAD+
-
Plasmodium falciparum
NADH
-
Plasmodium falciparum