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Literature summary 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)

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

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.40 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
1.1.1.40 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
1.1.1.40 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
1.1.1.40 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
1.1.1.40 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
1.1.1.40 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
1.1.1.40 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
1.1.1.40 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

EC Number Inhibitors Comment Organism Structure
1.1.1.40 CuCl2 about 40% loss of activity within 60 min Zea mays
1.1.1.40 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
1.1.1.40 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
1.1.1.40 additional information ZmnonC4-NADP-ME activity is not significantly modified by any chemical oxidant in 60 min Zea mays

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.40 0.00014
-
 (S)-malate pH 8.0, 30°C, mutant C246A Zea mays
1.1.1.40 0.00015
-
 (S)-malate pH 7.0, 30°C, wild-type, treated with dithiothreitol Zea mays
1.1.1.40 0.00017
-
 (S)-malate pH 8.0, 30°C, wild-type, treated with diamide Zea mays
1.1.1.40 0.00039
-
 (S)-malate pH 8.0, 30°C, wild-type, treated with dithiothreitol Zea mays
1.1.1.40 0.0008
-
 (S)-malate pH 7.0, 30°C, wild-type, treated with diamide Zea mays
1.1.1.40 0.0017
-
 (S)-malate pH 8.0, 30°C, mutant C231A Zea mays
1.1.1.40 0.003
-
 (S)-malate pH 8.0, 30°C, mutant C192A Zea mays
1.1.1.40 0.0036
-
 NADP+ pH 7.0, 30°C, oxidized isozyme ZmC4-NADP-ME Zea mays
1.1.1.40 0.0036
-
 NADP+ pH 7.0, 30°C, wild-type, treated with diamide Zea mays
1.1.1.40 0.0043
-
 (S)-malate pH 8.0, 30°C, mutant C270A Zea mays
1.1.1.40 0.0053
-
 NADP+ pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C246A Zea mays
1.1.1.40 0.0053
-
 NADP+ pH 8.0, 30°C, mutant C246A Zea mays
1.1.1.40 0.0088
-
 NADP+ pH 8.0, 30°C, oxidized isozyme ZmC4-NADP-ME Zea mays
1.1.1.40 0.0088
-
 NADP+ pH 8.0, 30°C, wild-type, treated with diamide Zea mays
1.1.1.40 0.0135
-
 NADP+ pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C270A Zea mays
1.1.1.40 0.0135
-
 NADP+ pH 8.0, 30°C, mutant C270A Zea mays
1.1.1.40 0.0147
-
 NADP+ pH 7.0, 30°C, reduced isozyme ZmC4-NADP-ME Zea mays
1.1.1.40 0.0147
-
 NADP+ pH 7.0, 30°C, wild-type, treated with dithiothreitol Zea mays
1.1.1.40 0.015
-
 NADP+ pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C231A Zea mays
1.1.1.40 0.015
-
 NADP+ pH 8.0, 30°C, mutant C231A Zea mays
1.1.1.40 0.0154
-
 NADP+ pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C192A Zea mays
1.1.1.40 0.0154
-
 NADP+ pH 8.0, 30°C, mutant C192A Zea mays
1.1.1.40 0.0276
-
 NADP+ pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME Zea mays
1.1.1.40 0.0276
-
 NADP+ pH 8.0, 30°C, wild-type, treated with dithiothreitol Zea mays
1.1.1.40 0.08
-
 (S)-malate pH 7.0, 30°C, oxidized isozyme ZmC4-NADP-ME Zea mays
1.1.1.40 0.14
-
 (S)-malate pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C246A Zea mays
1.1.1.40 0.15
-
 (S)-malate pH 7.0, 30°C, reduced isozyme ZmC4-NADP-ME Zea mays
1.1.1.40 0.17
-
 (S)-malate pH 8.0, 30°C, oxidized isozyme ZmC4-NADP-ME Zea mays
1.1.1.40 0.39
-
 (S)-malate pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME Zea mays
1.1.1.40 1.7
-
 (S)-malate pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C231A Zea mays
1.1.1.40 3
-
 (S)-malate pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C192A Zea mays
1.1.1.40 4.3
-
 (S)-malate pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C270A Zea mays

Localization

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

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.1.1.40 Mg2+ required Zea mays

Natural Substrates/ Products (Substrates)

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

Organism

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

Purification (Commentary)

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

Renatured (Commentary)

EC Number Renatured (Comment) Organism
1.1.1.40 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

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

Substrates and Products (Substrate)

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

Subunits

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

Synonyms

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

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.40 30
-
 assay at Zea mays

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.40 1.1
-
 NADP+ pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C246A Zea mays
1.1.1.40 1.1
-
 (S)-malate pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C246A Zea mays
1.1.1.40 1.1
-
 NADP+ pH 8.0, 30°C, mutant C246A Zea mays
1.1.1.40 1.6
-
 NADP+ pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C270A Zea mays
1.1.1.40 1.6
-
 (S)-malate pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C270A Zea mays
1.1.1.40 1.6
-
 NADP+ pH 8.0, 30°C, mutant C270A Zea mays
1.1.1.40 3.55
-
 NADP+ pH 7.0, 30°C, oxidized isozyme ZmC4-NADP-ME Zea mays
1.1.1.40 3.55
-
 (S)-malate pH 7.0, 30°C, oxidized isozyme ZmC4-NADP-ME Zea mays
1.1.1.40 3.55
-
 NADP+ pH 7.0, 30°C, wild-type, treated with diamide Zea mays
1.1.1.40 5.6
-
 NADP+ pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C192A Zea mays
1.1.1.40 5.6
-
 (S)-malate pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C192A Zea mays
1.1.1.40 5.6
-
 NADP+ pH 8.0, 30°C, mutant C192A Zea mays
1.1.1.40 8.35
-
 NADP+ pH 7.0, 30°C, reduced isozyme ZmC4-NADP-ME Zea mays
1.1.1.40 8.35
-
 (S)-malate pH 7.0, 30°C, reduced isozyme ZmC4-NADP-ME Zea mays
1.1.1.40 8.35
-
 NADP+ pH 7.0, 30°C, wild-type, treated with dithiothreitol Zea mays
1.1.1.40 23.2
-
 NADP+ pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C231A Zea mays
1.1.1.40 23.2
-
 (S)-malate pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C231A Zea mays
1.1.1.40 23.2
-
 NADP+ pH 8.0, 30°C, mutant C231A Zea mays
1.1.1.40 91
-
 NADP+ pH 8.0, 30°C, oxidized isozyme ZmC4-NADP-ME Zea mays
1.1.1.40 91
-
 (S)-malate pH 8.0, 30°C, oxidized isozyme ZmC4-NADP-ME Zea mays
1.1.1.40 91
-
 NADP+ pH 8.0, 30°C, wild-type, treated with diamide Zea mays
1.1.1.40 269
-
 NADP+ pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME Zea mays
1.1.1.40 269
-
 (S)-malate pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME Zea mays
1.1.1.40 269
-
 NADP+ pH 8.0, 30°C, wild-type, treated with dithiothreitol Zea mays

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.40 7 8 assay at Zea mays

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.40 NADP+
-
 Zea mays

General Information

EC Number General Information Comment Organism
1.1.1.40 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
1.1.1.40 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
1.1.1.40 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]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.1.40 0.12
-
 NADP+ pH 8.0, 30°C, mutant C270A Zea mays
1.1.1.40 0.14
-
 (S)-malate pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C246A Zea mays
1.1.1.40 0.21
-
 NADP+ pH 8.0, 30°C, mutant C246A Zea mays
1.1.1.40 0.36
-
 NADP+ pH 8.0, 30°C, mutant C192A Zea mays
1.1.1.40 0.37
-
 NADP+ pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C270A Zea mays
1.1.1.40 0.37
-
 (S)-malate pH 8.0, 30°C, mutant C270A Zea mays
1.1.1.40 1.5
-
 NADP+ pH 8.0, 30°C, mutant C231A Zea mays
1.1.1.40 1.7
-
 (S)-malate pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C231A Zea mays
1.1.1.40 1.86
-
 NADP+ pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C192A Zea mays
1.1.1.40 1.9
-
 (S)-malate pH 8.0, 30°C, mutant C192A Zea mays
1.1.1.40 3
-
 (S)-malate pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C192A Zea mays
1.1.1.40 4.3
-
 (S)-malate pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C270A Zea mays
1.1.1.40 7.8
-
 (S)-malate pH 8.0, 30°C, mutant C246A Zea mays
1.1.1.40 7.86
-
 NADP+ pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C246A Zea mays
1.1.1.40 9.7
-
 NADP+ pH 8.0, 30°C, wild-type, treated with dithiothreitol Zea mays
1.1.1.40 13.6
-
 (S)-malate pH 8.0, 30°C, mutant C231A Zea mays
1.1.1.40 13.64
-
 NADP+ pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME, mutant C231A Zea mays
1.1.1.40 689.7
-
 (S)-malate pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME Zea mays
1.1.1.40 690
-
 (S)-malate pH 8.0, 30°C, wild-type, treated with dithiothreitol Zea mays
1.1.1.40 9746
-
 NADP+ pH 8.0, 30°C, reduced isozyme ZmC4-NADP-ME Zea mays