Cloned (Comment) | Organism |
---|---|
overexpression of His-tagged wild-type and mutants in Escherichia coli strain DH5alpha, functional complementation of the enzyme deficient Escherichia coli triple mutant strain EJ1321 | Zea mays |
Protein Variants | Comment | Organism |
---|---|---|
A387G | site directed mutagenesis, mutation at the NADP+ binding site, mutant shows 48fold decreased kcat and 4.3 and 5.8fold increased Km for NADP+ and L-malate, respectively, compared to the wild-type enzyme, no activity with NAD+ | Zea mays |
A392G | site directed mutagenesis, mutation at the NADP+ binding site, mutant shows unaltered kcat, but 3.5 and 2.6fold increased Km for NADP+ and L-malate, respectively, and increased activity with NAD+ compared to the wild-type enzyme | Zea mays |
additional information | construction of the Gsite5V and Gsite2V mutants, mutation at the NADP+ binding site | Zea mays |
R237L | site directed mutagenesis, mutation at the NADP+ binding site, mutant shows 530fold decreased kcat and 36.3 and 15.3fold increased Km for NADP+ and L-malate, respectively, compared to the wild-type enzyme, no activity with NAD+ | Zea mays |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
L-malate | pH 7.0, high concentration | Zea mays |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.008 | - |
NADP+ | wild-type enzyme, pH 7.0 | Zea mays | |
0.0086 | - |
NADP+ | recombinant enzyme, pH 7.0 | Zea mays | |
0.03 | - |
NADP+ | mutant A392G, pH 7.0 | Zea mays | |
0.037 | - |
NADP+ | mutant A387G, pH 7.0 | Zea mays | |
0.19 | - |
L-malate | recombinant enzyme, pH 7.0 | Zea mays | |
0.23 | - |
L-malate | wild-type enzyme, pH 7.0 | Zea mays | |
0.29 | - |
NADP+ | mutant R237L, pH 7.0 | Zea mays | |
0.5 | - |
L-malate | mutant A392G, pH 7.0 | Zea mays | |
1.1 | - |
L-malate | mutant A387G, pH 7.0 | Zea mays | |
2.9 | - |
L-malate | mutant R237L, pH 7.0 | Zea mays | |
6 | - |
NAD+ | mutant A392G, pH 7.0 | Zea mays | |
8.1 | - |
NAD+ | wild-type enzyme, pH 7.0 | Zea mays |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
chloroplast | - |
Zea mays | 9507 | - |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Zea mays |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
62000 | - |
x * 62000, detagged recombinant wild-type enzyme | Zea mays |
110000 | - |
recombinant dimeric enzyme, gel filtration at pH 7.0, minor peak | Zea mays |
226000 | - |
recombinant tetrameric enzyme, gel filtration at pH 8.0, major peak | Zea mays |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-malate + NAD(P)+ | Zea mays | the unique and specialized C4-type enzyme has evolved fro the C3-type enzyme | pyruvate + CO2 + NAD(P)H | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Zea mays | P16243 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged mutant enzyme to homogeneity from Escherichia coli by metal affinity chromatography | Zea mays |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
leaf | - |
Zea mays | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-malate + NAD(P)+ | - |
Zea mays | pyruvate + CO2 + NAD(P)H | - |
? | |
(S)-malate + NAD(P)+ | the unique and specialized C4-type enzyme has evolved fro the C3-type enzyme | Zea mays | pyruvate + CO2 + NAD(P)H | - |
? | |
(S)-malate + NAD+ | - |
Zea mays | pyruvate + CO2 + NADH | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | extent of oligomerization is pH-dependent, 3D-structure analysis | Zea mays |
oligomer | x * 62000, detagged recombinant wild-type enzyme | Zea mays |
Synonyms | Comment | Organism |
---|---|---|
NADP-malic enzyme | - |
Zea mays |
NADP-ME | - |
Zea mays |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.38 | - |
NADP+ | mutant R237L, pH 7.0 | Zea mays | |
4.2 | - |
NADP+ | mutant A387G, pH 7.0 | Zea mays | |
13.5 | - |
NAD+ | wild-type enzyme, pH 7.0 | Zea mays | |
30.8 | - |
NADP+ | recombinant enzyme, pH 7.0 | Zea mays | |
40.6 | - |
NAD+ | mutant A392G, pH 7.0 | Zea mays | |
200.3 | - |
NADP+ | mutant A392G, pH 7.0 | Zea mays | |
201.3 | - |
NADP+ | wild-type enzyme, pH 7.0 | Zea mays |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
recombinant enzyme | Zea mays |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | the recombinant wild-type enzyme shows a low intrinsic activity with NAD+, best at pH 7.0, mutant A392G shows increased activity with NAD+ compared to the wild-type enzyme | Zea mays | |
NADP+ | binding site structure, Arg237 is important for cofactor binding and specificity | Zea mays |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
4.7 | - |
L-malate | recombinant enzyme, pH 7.0 | Zea mays |
Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|
Zea mays | recombinant enzyme, isoelectric focusing | - |
7.1 |