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Literature summary for 1.1.1.37 extracted from
- Purification and characterization of the plastid-localized NAD-dependent malate dehydrogenase from Arabidopsis thaliana (2016), Biotechnol. Appl. Biochem., 63, 490-496.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene mdh, sequence comparisons, recombinant expression of N-terminally His-Sumo-tagged enzyme in Escherichia coli stain BL21 (DE3) | Arabidopsis thaliana |
Crystallization (Commentary)
| Crystallization (Comment) | Organism |
|---|---|
| purified recombinant detagged enzyme, hanging drop vapor diffusion method, mixing of 0.0003 ml 6 mg/ml protein in 20 mM Tris-HCl, pH 8.0, 200 mM NaCl, and 10% glycerol, with 0.0003 ml of reservoir solution containing 1.2 M sodium citrate tribasic dihydrate, pH 7.0, 20°C, method optimization, X-ray diffraction structure determination and analysis | Arabidopsis thaliana |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| Cu2+ | complete inhibition at 5 mM | Arabidopsis thaliana |  |
| Fe3+ | complete inhibition at 5 mM | Arabidopsis thaliana | |
| Zn2+ | 42% inhibition at 5 mM | Arabidopsis thaliana | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | Michaelis-Menten kinetics | Arabidopsis thaliana | |
| 0.324 | - |
oxaloacetate | pH 7.5, 30°C, recombinant enzyme | Arabidopsis thaliana | |
| 0.431 | - |
NADH | pH 7.5, 30°C, recombinant enzyme | Arabidopsis thaliana | |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| chloroplast | the enzyme harbors a transit peptide of 80 amino acids at the N-terminus | Arabidopsis thaliana | 9507 | - |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Mn2+ | 24% activation at 5 mM | Arabidopsis thaliana | |
| additional information | poor effect by 5 mM of Mg2+ or Ca2+ | Arabidopsis thaliana |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 36500 | - |
- |
Arabidopsis thaliana |
| 45600 | - |
- |
Arabidopsis thaliana |
| 67000 | - |
about, recombinant detagged mature enzyme, gel filtration | Arabidopsis thaliana |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| (S)-malate + NAD+ | Arabidopsis thaliana | - |
oxaloacetate + NADH + H+ | - |
r | |
| oxaloacetate + NADH + H+ | Arabidopsis thaliana | - |
(S)-malate + NAD+ | - |
r | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Arabidopsis thaliana | Q9SN86 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-Sumo-tagged enzyme from Escherichia coli stain BL21 (DE3) by nickel affinity chromatography, proteolytic cleavage of the His-Sumo tag, ultrafiltration, and gel filtration | Arabidopsis thaliana |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| (S)-malate + NAD+ | - |
Arabidopsis thaliana | oxaloacetate + NADH + H+ | - |
r | |
| oxaloacetate + NADH + H+ | - |
Arabidopsis thaliana | (S)-malate + NAD+ | - |
r | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| homodimer | 2 * 45600, full-length enzyme, SDS-PAGE, 2 * 36500, mature enzyme without transit peptide, SDS-PAGE and glutaraldehyde cross-linking of plNAD-MDH | Arabidopsis thaliana |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| MDH | - |
Arabidopsis thaliana |
| NAD-dependent malate dehydrogenase | - |
Arabidopsis thaliana |
| plastid-localized NAD-dependent MDH | - |
Arabidopsis thaliana |
| plNAD-MDH | - |
Arabidopsis thaliana |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | - |
assay at | Arabidopsis thaliana |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.5 | - |
- |
Arabidopsis thaliana |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 5 | 10 | maximal activity at pH 7.5, 70% of maximal activity at pH 10.0, 40% at pH 7.0, 20% at pH 5.0, very low activity below pH 5.0 and above pH 10.0 | Arabidopsis thaliana |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| NAD+ | - |
Arabidopsis thaliana | |
| NADH | - |
Arabidopsis thaliana | |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | Arabidopsis thaliana contains 10 MDHs with only one single copy of MDH gene in the chloroplast, which is a plastidlocalized NAD-dependent MDH | Arabidopsis thaliana |
| physiological function | the plastid-localized NAD-dependent MDH is important for plant survival in a dark or shady environment under which plNAD-MDH replaces the inactive chloroplast NADP-MDH in the regeneration of NAD+ to produce ATP | Arabidopsis thaliana |
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