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Literature summary for 1.8.1.4 extracted from
- The pyruvate and alpha-ketoglutarate dehydrogenase complexes of Pseudomonas aeruginosa catalyze pyocyanin and phenazine-1-carboxylic acid reduction via the subunit dihydrolipoamide dehydrogenase (2017), J. Biol. Chem., 292, 5593-5607 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene lpd3, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) | Pseudomonas aeruginosa |
| gene lpdG, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Pseudomonas aeruginosa |
| gene lpdV, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) | Pseudomonas aeruginosa |
Crystallization (Commentary)
| Crystallization (Comment) | Organism |
|---|---|
| purified enzyme in apoform, and in complexes with Nad+ and NADH, X-ray diffraction structure determination and analysis at 1.35-1.79 A resolution | Pseudomonas aeruginosa |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| I192G | site-directed mutagenesis, the mutant is active with phenazine-1-carboxylic acid | Pseudomonas aeruginosa |
| V191Y | site-directed mutagenesis, the mutant is active with phenazine-1-carboxylic acid | Pseudomonas aeruginosa |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| cyanide | slight inhibition; slight inhibition; slight inhibition | Pseudomonas aeruginosa |  |
| diphenyleneiodonium | an inhibitor of flavoproteins and heme-containing proteins, effectively inhibits phenazine reduction in vitro; an inhibitor of flavoproteins and heme-containing proteins, effectively inhibits phenazine reduction in vitro; an inhibitor of flavoproteins and heme-containing proteins, effectively inhibits phenazine reduction in vitro | Pseudomonas aeruginosa | |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| dihydrolipoamide + NAD+ | Pseudomonas aeruginosa | - |
lipoamide + NADH + H+ | - |
r | |
| dihydrolipoamide + NAD+ | Pseudomonas aeruginosa UCBPP-PA14 | - |
lipoamide + NADH + H+ | - |
r | |
| phenazine-1-carboxylic acid + NADH + H+ | Pseudomonas aeruginosa | by cell lysate | reduced phenazine-1-carboxylic acid + NAD+ | - |
? | |
| phenazine-1-carboxylic acid + NADH + H+ | Pseudomonas aeruginosa UCBPP-PA14 | by cell lysate | reduced phenazine-1-carboxylic acid + NAD+ | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Pseudomonas aeruginosa | A0A0H2Z9F5 | - |
- |
| Pseudomonas aeruginosa | A0A0H2ZB32 | - |
- |
| Pseudomonas aeruginosa | A0A0H2ZHZ0 | - |
- |
| Pseudomonas aeruginosa UCBPP-PA14 | A0A0H2Z9F5 | - |
- |
| Pseudomonas aeruginosa UCBPP-PA14 | A0A0H2ZB32 | - |
- |
| Pseudomonas aeruginosa UCBPP-PA14 | A0A0H2ZHZ0 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, ultrafiltration, and gel filtration | Pseudomonas aeruginosa |
| recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, ultrafiltration, and gel filtration. Purification of native pyruvate and2-oxoglutarate dehydrogenase complexes from strain PA14 by hydroxyapatite chromatography and gel filtration | Pseudomonas aeruginosa |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| dihydrolipoamide + NAD+ | - |
Pseudomonas aeruginosa | lipoamide + NADH + H+ | - |
r | |
| dihydrolipoamide + NAD+ | - |
Pseudomonas aeruginosa UCBPP-PA14 | lipoamide + NADH + H+ | - |
r | |
| methylene blue + NADH + H+ | - |
Pseudomonas aeruginosa | reduced methylene blue + NAD+ | - |
? | |
| methylene blue + NADH + H+ | - |
Pseudomonas aeruginosa UCBPP-PA14 | reduced methylene blue + NAD+ | - |
? | |
| additional information | phenazines may substitute for NAD+ in LpdG and other enzymes, achieving the same end by a different mechanism. PCA and pyocyanin reduction by the purified complexes required all substrates and cofactors | Pseudomonas aeruginosa | ? | - |
? | |
| additional information | phenazines may substitute for NAD+ in LpdG and other enzymes, achieving the same end by a different mechanism. PCA and pyocyanin reduction by the purified complexes required all substrates and cofactors | Pseudomonas aeruginosa UCBPP-PA14 | ? | - |
? | |
| phenazine-1-carboxylic acid + NADH + H+ | by cell lysate | Pseudomonas aeruginosa | reduced phenazine-1-carboxylic acid + NAD+ | - |
? | |
| phenazine-1-carboxylic acid + NADH + H+ | PCA, the precursor for all biological phenazines in Pseudomonas aeruginosa, promotes anaerobic energy generation by redox cycling. Enzyme LpdG residues Val191 and Ile192 do not sterically hinder PCA frombinding to LpdG | Pseudomonas aeruginosa | reduced phenazine-1-carboxylic acid + NAD+ | - |
? | |
| phenazine-1-carboxylic acid + NADH + H+ | the precursor for all biological phenazines in Pseudomonas aeruginosa, promotes anaerobic energy generation by redox cycling | Pseudomonas aeruginosa | reduced phenazine-1-carboxylic acid + NAD+ | - |
? | |
| phenazine-1-carboxylic acid + NADH + H+ | by cell lysate | Pseudomonas aeruginosa UCBPP-PA14 | reduced phenazine-1-carboxylic acid + NAD+ | - |
? | |
| phenazine-1-carboxylic acid + NADH + H+ | PCA, the precursor for all biological phenazines in Pseudomonas aeruginosa, promotes anaerobic energy generation by redox cycling. Enzyme LpdG residues Val191 and Ile192 do not sterically hinder PCA frombinding to LpdG | Pseudomonas aeruginosa UCBPP-PA14 | reduced phenazine-1-carboxylic acid + NAD+ | - |
? | |
| phenazine-1-carboxylic acid + NADH + H+ | the precursor for all biological phenazines in Pseudomonas aeruginosa, promotes anaerobic energy generation by redox cycling | Pseudomonas aeruginosa UCBPP-PA14 | reduced phenazine-1-carboxylic acid + NAD+ | - |
? | |
| pyocyanin + NADH + H+ | pyocyanin is reported to stimulate respiration | Pseudomonas aeruginosa | reduced pyocyanin + NAD+ | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| DLDH | - |
Pseudomonas aeruginosa |
| Lpd3 | - |
Pseudomonas aeruginosa |
| LpdG | - |
Pseudomonas aeruginosa |
| LpdV | - |
Pseudomonas aeruginosa |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 28 | - |
assay at | Pseudomonas aeruginosa |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.5 | - |
assay at | Pseudomonas aeruginosa |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| FAD | - |
Pseudomonas aeruginosa | |
| FAD | binding structure analysis | Pseudomonas aeruginosa | |
| NAD+ | - |
Pseudomonas aeruginosa | |
| NAD+ | binding structure analysis | Pseudomonas aeruginosa | |
| NADH | - |
Pseudomonas aeruginosa | |
| NADH | binding structure analysis | Pseudomonas aeruginosa | |
General Information
| General Information | Comment | Organism |
|---|---|---|
| additional information | purification of an NADH:PCA or NADPH:PCA oxidoreductase, active with phenazine-1-carboxylic acid and other phenazines, from Pseudomonas aeruginosa cell lysate is not successful | Pseudomonas aeruginosa |
| additional information | purification of an NADH:PCA or NADPH:PCA oxidoreductase, active with phenazine-1-carboxylic acid and other phenazines, from Pseudomonas aeruginosa cell lysate is not successful. Structural analysis of LpdG, overview | Pseudomonas aeruginosa |
| physiological function | LpdG, not LpdV and Lpd3, is the primary DLDH of the Pseudomonas aeruginosa PA14 pyruvate dehydrogenase, and is the enzymatically relevant DLDH for both pyruvate and 2-oxoglutarate dehydrogenase | Pseudomonas aeruginosa |
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Release 2023.1 (January 2023)
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