Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 1.1.1.40 extracted from

  • Alvarez, C.; Detarsio, E.; Moreno, S.; Andreo, C.; Drincovich, M.
    Functional characterization of residues involved in redox modulation of maize photosynthetic NADP-malic enzyme activity (2012), Plant Cell Physiol., 53, 1144-1153.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Zea mays

Protein Variants

Protein Variants Comment Organism
C192A site-directed mutagenesis of isozyme ZmC4-NADP-ME, the mutation does not affect the tetrameric state, the mutant displays lower malate affinity than the wild-type enzyme Zea mays
C192A marked decrease in kcat value, less than 10% of wild-type, with concomitant increase in Km value for NADP+. Unlike wild-type, activity is not significantly changed in presence of oxidant iodosobenzoate Zea mays
C231A site-directed mutagenesis of isozyme ZmC4-NADP-ME, the mutation does not affect the tetrameric state, the mutant displays lower malate affinity than the wild-type enzyme Zea mays
C231A marked decrease in kcat value, less than 10% of wild-type, with concomitant increase in Km value for NADP+. Similar to wild-type, activity decreases in presence of oxidant iodosobenzoate Zea mays
C246A site-directed mutagenesis of isozyme ZmC4-NADP-ME, the mutation does not affect the tetrameric state, C246A exhibits a nearly 5fold increase in its affinity towards NADP+ and a 3fold decrease for malate compared to the wild-tpe enzyme Zea mays
C246A marked decrease in kcat value, less than 10% of wild-type, with concomitant increase in Km value for NADP+. Unlike wild-type, activity is not significantly changed in presence of oxidant iodosobenzoate Zea mays
C270A site-directed mutagenesis of isozyme ZmC4-NADP-ME, the mutation does not affect the tetrameric state, the mutant displays lower malate affinity than the wild-type enzyme Zea mays
C270A marked decrease in kcat value, less than 10% of wild-type, with concomitant increase in Km value for NADP+. Unlike wild-type, activity is not significantly changed in presence of oxidant iodosobenzoate Zea mays

Inhibitors

Inhibitors Comment Organism Structure
CuCl2 about 40% loss of activity within 60 min Zea mays
Diamide 2 mM, time-dependent decrease in activity reaching about 40% of initial activity after 60 min. In presence of dithiothreitol, a complete recovery is observed after 90 min. Enzyme oxidation decreases the catalytic activity. No severe loss of protein secondary structure takes place after oxidation; about 40% loss of activity within 60 min Zea mays
Iodosobenzoate 1 mM, time-dependent decrease in activity reaching about 40% of initial activity after 60 min; about 40% loss of activity within 60 min Zea mays
additional information ZmnonC4-NADP-ME activity is not significantly modified by any chemical oxidant in 60 min Zea mays

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.00014
-
(S)-malate pH 8.0, 30°C, mutant C246A Zea mays
0.00015
-
(S)-malate pH 7.0, 30°C, wild-type, treated with dithiothreitol Zea mays
0.00017
-
(S)-malate pH 8.0, 30°C, wild-type, treated with diamide Zea mays
0.00039
-
(S)-malate pH 8.0, 30°C, wild-type, treated with dithiothreitol Zea mays
0.0008
-
(S)-malate pH 7.0, 30°C, wild-type, treated with diamide Zea mays
0.0017
-
(S)-malate pH 8.0, 30°C, mutant C231A Zea mays
0.003
-
(S)-malate pH 8.0, 30°C, mutant C192A Zea mays
0.0036
-
NADP+ pH 7.0, 30°C, oxidized isozyme ZmC4-NADP-ME Zea mays
0.0036
-
NADP+ pH 7.0, 30°C, wild-type, treated with diamide Zea mays
0.0043
-
(S)-malate pH 8.0, 30°C, mutant C270A Zea mays
0.0053
-
NADP+ pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C246A Zea mays
0.0053
-
NADP+ pH 8.0, 30°C, mutant C246A Zea mays
0.0088
-
NADP+ pH 8.0, 30°C, oxidized isozyme ZmC4-NADP-ME Zea mays
0.0088
-
NADP+ pH 8.0, 30°C, wild-type, treated with diamide Zea mays
0.0135
-
NADP+ pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C270A Zea mays
0.0135
-
NADP+ pH 8.0, 30°C, mutant C270A Zea mays
0.0147
-
NADP+ pH 7.0, 30°C, reduced isozyme ZmC4-NADP-ME Zea mays
0.0147
-
NADP+ pH 7.0, 30°C, wild-type, treated with dithiothreitol Zea mays
0.015
-
NADP+ pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C231A Zea mays
0.015
-
NADP+ pH 8.0, 30°C, mutant C231A Zea mays
0.0154
-
NADP+ pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C192A Zea mays
0.0154
-
NADP+ pH 8.0, 30°C, mutant C192A Zea mays
0.0276
-
NADP+ pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME Zea mays
0.0276
-
NADP+ pH 8.0, 30°C, wild-type, treated with dithiothreitol Zea mays
0.08
-
(S)-malate pH 7.0, 30°C, oxidized isozyme ZmC4-NADP-ME Zea mays
0.14
-
(S)-malate pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C246A Zea mays
0.15
-
(S)-malate pH 7.0, 30°C, reduced isozyme ZmC4-NADP-ME Zea mays
0.17
-
(S)-malate pH 8.0, 30°C, oxidized isozyme ZmC4-NADP-ME Zea mays
0.39
-
(S)-malate pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME Zea mays
1.7
-
(S)-malate pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C231A Zea mays
3
-
(S)-malate pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C192A Zea mays
4.3
-
(S)-malate pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C270A Zea mays

Localization

Localization Comment Organism GeneOntology No. Textmining
chloroplast isozymes ZmC4-NADP-ME and ZmnonC4-NADP-ME Zea mays 9507
-
plastid
-
Zea mays 9536
-

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Zea mays

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
(S)-malate + NADP+ Zea mays
-
pyruvate + CO2 + NADPH
-
?

Organism

Organism UniProt Comment Textmining
Zea mays
-
isozymes ZmC4-NADP-ME and ZmnonC4-NADP-ME
-
Zea mays P16243
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) to homogeneity Zea mays

Renatured (Commentary)

Renatured (Comment) Organism
presence of 2 mM diamide leads to a time-dependent decrease in activity, reaching about 40% of initial activity after 60 min. After addition of dithiothreitol, a complete recovery is observed after 90 min Zea mays

Source Tissue

Source Tissue Comment Organism Textmining
bundle sheath cell isozyme ZmC4-NADP-ME Zea mays
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(S)-malate + NADP+
-
Zea mays pyruvate + CO2 + NADPH
-
?

Subunits

Subunits Comment Organism
tetramer recombinant ZmC4-NADP-ME wild-type and mutants Zea mays

Synonyms

Synonyms Comment Organism
C4-NADP-ME isoform involved in C4 photosynthesis Zea mays
NADP-malic enzyme
-
Zea mays
NADP-ME
-
Zea mays

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Zea mays

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1
-
NADP+ pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C246A Zea mays
1.1
-
(S)-malate pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C246A Zea mays
1.1
-
NADP+ pH 8.0, 30°C, mutant C246A Zea mays
1.6
-
NADP+ pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C270A Zea mays
1.6
-
(S)-malate pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C270A Zea mays
1.6
-
NADP+ pH 8.0, 30°C, mutant C270A Zea mays
3.55
-
NADP+ pH 7.0, 30°C, oxidized isozyme ZmC4-NADP-ME Zea mays
3.55
-
(S)-malate pH 7.0, 30°C, oxidized isozyme ZmC4-NADP-ME Zea mays
3.55
-
NADP+ pH 7.0, 30°C, wild-type, treated with diamide Zea mays
5.6
-
NADP+ pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C192A Zea mays
5.6
-
(S)-malate pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C192A Zea mays
5.6
-
NADP+ pH 8.0, 30°C, mutant C192A Zea mays
8.35
-
NADP+ pH 7.0, 30°C, reduced isozyme ZmC4-NADP-ME Zea mays
8.35
-
(S)-malate pH 7.0, 30°C, reduced isozyme ZmC4-NADP-ME Zea mays
8.35
-
NADP+ pH 7.0, 30°C, wild-type, treated with dithiothreitol Zea mays
23.2
-
NADP+ pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C231A Zea mays
23.2
-
(S)-malate pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C231A Zea mays
23.2
-
NADP+ pH 8.0, 30°C, mutant C231A Zea mays
91
-
NADP+ pH 8.0, 30°C, oxidized isozyme ZmC4-NADP-ME Zea mays
91
-
(S)-malate pH 8.0, 30°C, oxidized isozyme ZmC4-NADP-ME Zea mays
91
-
NADP+ pH 8.0, 30°C, wild-type, treated with diamide Zea mays
269
-
NADP+ pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME Zea mays
269
-
(S)-malate pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME Zea mays
269
-
NADP+ pH 8.0, 30°C, wild-type, treated with dithiothreitol Zea mays

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7 8 assay at Zea mays

Cofactor

Cofactor Comment Organism Structure
NADP+
-
Zea mays

General Information

General Information Comment Organism
metabolism isoform C4-NADP-ME involved in C4 photosynthesis is modulated by redox status, and its oxidation produces a conformational change limiting the catalytic process, although inducing higher affinity binding of the substrates. Residues Cys192, Cys246, Cys270 and Cys410 are directly or indirectly implicated in C4-NADP-ME redox modulation Zea mays
additional information reversal of ZmC4-NADP-ME oxidation by chemical reductants, e.g. iodosobenzoate and CuCl2, doe to the presence of thiol groups able to form disulfide bonds. Residues Cys192, Cys246, Cys270, and Cys410 may be directly or indirectly implicated in ZmC4-NADP-ME redox modulation. Redox regulation plays a key role in many plastid functions. Isozyme specific redox regulation of ZmC4-NADP-ME activity, the modulation is not observed in the case of isozyme ZmnonC4-NADP-ME. The replacement of Cys246 with serine in ZmnonC4-NADP-ME may be responsible for the absence of redox modulation Zea mays
physiological function one isozyme exclusively expressed in the bundle sheath cells and involved in C4 photosynthesis, i.e. ZmC4-NADP-ME, and the other, ZmnonC4-NADP-ME, with housekeeping roles Zea mays

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.12
-
NADP+ pH 8.0, 30°C, mutant C270A Zea mays
0.14
-
(S)-malate pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C246A Zea mays
0.21
-
NADP+ pH 8.0, 30°C, mutant C246A Zea mays
0.36
-
NADP+ pH 8.0, 30°C, mutant C192A Zea mays
0.37
-
NADP+ pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C270A Zea mays
0.37
-
(S)-malate pH 8.0, 30°C, mutant C270A Zea mays
1.5
-
NADP+ pH 8.0, 30°C, mutant C231A Zea mays
1.7
-
(S)-malate pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C231A Zea mays
1.86
-
NADP+ pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C192A Zea mays
1.9
-
(S)-malate pH 8.0, 30°C, mutant C192A Zea mays
3
-
(S)-malate pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C192A Zea mays
4.3
-
(S)-malate pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C270A Zea mays
7.8
-
(S)-malate pH 8.0, 30°C, mutant C246A Zea mays
7.86
-
NADP+ pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C246A Zea mays
9.7
-
NADP+ pH 8.0, 30°C, wild-type, treated with dithiothreitol Zea mays
13.6
-
(S)-malate pH 8.0, 30°C, mutant C231A Zea mays
13.64
-
NADP+ pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C231A Zea mays
689.7
-
(S)-malate pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME Zea mays
690
-
(S)-malate pH 8.0, 30°C, wild-type, treated with dithiothreitol Zea mays
9746
-
NADP+ pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME Zea mays