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Literature summary for 1.1.1.40 extracted from

  • Leroux, A.; Maugeri, D.; Opperdoes, F.; Cazzulo, J.; Nowicki, C.
    Comparative studies on the biochemical properties of the malic enzymes from Trypanosoma cruzi and Trypanosoma brucei (2011), FEMS Microbiol. Lett., 314, 25-33.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
L-aspartate 10fold activation Trypanosoma cruzi
L-aspartate the cytosolic isozyme is 50% activated by L-aspartate Trypanosoma brucei
L-aspartate the cytosolic isozyme is over 10fold activated by L-aspartate Trypanosoma cruzi

Cloned(Commentary)

Cloned (Comment) Organism
cytosolic and mitochondrial isozymes, expression in of His-tagged isozymes ME1 and ME2 Escherichia coli Rosetta (DE3)pLysS Trypanosoma brucei
cytosolic and mitochondrial isozymes, expression of His-tagged ME2 in Escherichia coli Rosetta (DE3)pLysS and of His-tagged ME1 in Escherichia coli BL(DE3) Trypanosoma cruzi
expression of His-tagged isozymes ME1 and ME2 in Escherichia coli strain BL21(DE3) Trypanosoma brucei
expression of His-tagged isozymes ME1 and ME2 in Escherichia coli strain BL21(DE3) Trypanosoma cruzi

Crystallization (Commentary)

Crystallization (Comment) Organism
recombinant His-tagged cytosolic and mitochondrial isozymes from Escherichia coli by nickel affinity chromatography Trypanosoma brucei

Inhibitors

Inhibitors Comment Organism Structure
oxaloacetate competitive Trypanosoma brucei
oxaloacetate competitive Trypanosoma cruzi

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.016
-
NADP+ isozyme ME2, pH 7.4, temperature not specified in the publication Trypanosoma brucei
0.016
-
NADP+ pH and temperature not specified in the publication, recombinant isozyme ME2, in presence of 0.5 mM Mn2+ Trypanosoma brucei
0.027
-
NADP+ isozyme ME1, pH 7.4, temperature not specified in the publication Trypanosoma cruzi
0.027
-
NADP+ pH and temperature not specified in the publication, recombinant isozyme ME1, in presence of 0.5 mM Mn2+ Trypanosoma cruzi
0.032
-
NADP+ isozyme ME2, pH 7.4, temperature not specified in the publication Trypanosoma cruzi
0.032
-
NADP+ pH and temperature not specified in the publication, recombinant isozyme ME2, in presence of 0.5 mM Mn2+ Trypanosoma cruzi
0.048
-
NADP+ isozyme ME1, pH 7.4, temperature not specified in the publication Trypanosoma brucei
0.048
-
NADP+ pH and temperature not specified in the publication, recombinant isozyme ME1, in presence of 0.5 mM Mn2+ Trypanosoma brucei
0.13
-
(S)-malate isozyme ME1, pH 7.4, temperature not specified in the publication Trypanosoma cruzi
0.13
-
(S)-malate pH and temperature not specified in the publication, recombinant isozyme ME1, in presence of 0.5 mM Mn2+ Trypanosoma cruzi
0.32
-
(S)-malate isozyme ME1, pH 7.4, temperature not specified in the publication Trypanosoma brucei
0.32
-
(S)-malate pH and temperature not specified in the publication, recombinant isozyme ME1, in presence of 0.5 mM Mn2+ Trypanosoma brucei
0.49
-
(S)-malate isozyme ME2, pH 7.4, temperature not specified in the publication Trypanosoma brucei
0.49
-
(S)-malate pH and temperature not specified in the publication, recombinant isozyme ME2, in presence of 0.5 mM Mn2+ Trypanosoma brucei
2.3
-
(S)-malate isozyme ME2, pH 7.4, temperature not specified in the publication Trypanosoma cruzi
2.3
-
(S)-malate pH and temperature not specified in the publication, recombinant isozyme ME2, in presence of 0.5 mM Mn2+ Trypanosoma cruzi
4.3
-
pyruvate isozyme ME1, with NADPH and CO2, pH 7.4, temperature not specified in the publication Trypanosoma brucei
4.3
-
pyruvate pH and temperature not specified in the publication, recombinant isozyme ME1, in presence of 0.5 mM Mn2+ Trypanosoma brucei
5.1
-
pyruvate isozyme ME2, with NADPH and CO2, pH 7.4, temperature not specified in the publication Trypanosoma cruzi
5.1
-
pyruvate pH and temperature not specified in the publication, recombinant isozyme ME2, in presence of 0.5 mM Mn2+ Trypanosoma cruzi
5.3
-
pyruvate isozyme ME2, with NADPH and CO2, pH 7.4, temperature not specified in the publication Trypanosoma brucei
5.3
-
pyruvate pH and temperature not specified in the publication, recombinant isozyme ME2, in presence of 0.5 mM Mn2+ Trypanosoma brucei
7.6
-
pyruvate isozyme ME1, with NADPH and CO2, pH 7.4, temperature not specified in the publication Trypanosoma cruzi
7.6
-
pyruvate pH and temperature not specified in the publication, recombinant isozyme ME1, in presence of 0.5 mM Mn2+ Trypanosoma cruzi

Localization

Localization Comment Organism GeneOntology No. Textmining
cytosol cytosolic isozyme, appears to be clearly more abundant in the insect stage, although it can be immunodetected in the bloodstream form Trypanosoma brucei 5829
-
cytosol cytosolic isozyme, the cytosolic isoform appears to be more abundant in the insect stages of the parasite Trypanosoma cruzi 5829
-
cytosol both cytosolic and mitochondrial isozymes are more abundant in the insect stage, although they can be immunodetected in the bloodstream forms Trypanosoma brucei 5829
-
cytosol the cytosolic isoform is more abundant in the insect stages of the parasite Trypanosoma cruzi 5829
-
mitochondrion mitochondrial isozyme, appears to be clearly more abundant in the insect stage, although it can be immunodetected in the bloodstream form Trypanosoma brucei 5739
-
mitochondrion mitochondrial isozyme, the expression of the mitochondrial isozyme seems to be clearly upregulated in amastigotes Trypanosoma cruzi 5739
-
mitochondrion both cytosolic and mitochondrial isozymes are more abundant in the insect stage, although they can be immunodetected in the bloodstream forms Trypanosoma brucei 5739
-
mitochondrion expression of the mitochondrial ME is upregulated in amastigotes Trypanosoma cruzi 5739
-

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ activates, kcat is 1280/s and Km 0.27 mM for ME1, and kcat is 208/s and Km 0.58 mM for ME2 Trypanosoma brucei
Mg2+ highly activating, kcat is 461/s and Km 0.04 mM for ME1, and kcat is 864/s and Km 0.57 mM for ME2 Trypanosoma cruzi
Mg2+ can partially substitute for Mn2+, activates Trypanosoma brucei
Mg2+ can partially substitute for Mn2+, activates Trypanosoma cruzi
Mn2+ activates, required Trypanosoma brucei
Mn2+ activates, required Trypanosoma cruzi
Mn2+ activates, kcat is 518/s and Km 0.0043 mM for ME1, and kcat is 687/s and Km 0.0091 mM for ME2 Trypanosoma cruzi
Mn2+ highly activating, kcat is 905/s and Km 0.0024 mM for ME1, and kcat is 246/s and Km 0.0066 mM for ME2 Trypanosoma brucei

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
(S)-malate + NADP+ Trypanosoma brucei
-
pyruvate + CO2 + NADPH
-
r
(S)-malate + NADP+ Trypanosoma cruzi
-
pyruvate + CO2 + NADPH
-
r
(S)-malate + NADP+ Trypanosoma brucei
-
pyruvate + NADPH + H+ + CO2
-
r
(S)-malate + NADP+ Trypanosoma cruzi
-
pyruvate + NADPH + H+ + CO2
-
?
(S)-malate + NADP+ Trypanosoma cruzi CL Brener
-
pyruvate + CO2 + NADPH
-
r
(S)-malate + NADP+ Trypanosoma cruzi CL Brener
-
pyruvate + NADPH + H+ + CO2
-
?
(S)-malate + NADP+ Trypanosoma brucei 427
-
pyruvate + CO2 + NADPH
-
r
(S)-malate + NADP+ Trypanosoma brucei 427
-
pyruvate + NADPH + H+ + CO2
-
r
(S)-malate + NADP+ Trypanosoma brucei stock 427
-
pyruvate + CO2 + NADPH
-
r
(S)-malate + NADP+ Trypanosoma brucei stock 427
-
pyruvate + NADPH + H+ + CO2
-
r

Organism

Organism UniProt Comment Textmining
Trypanosoma brucei
-
-
-
Trypanosoma brucei
-
grown in male Wistar rats
-
Trypanosoma brucei 427
-
-
-
Trypanosoma brucei stock 427
-
grown in male Wistar rats
-
Trypanosoma cruzi
-
-
-
Trypanosoma cruzi CL Brener
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged cytosolic and mitochondrial isozymes from Escherichia coli by nickel affinity chromatography Trypanosoma cruzi
recombinant His-tagged isozymes ME1 and ME2 from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Trypanosoma brucei
recombinant His-tagged isozymes ME1 and ME2 from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Trypanosoma cruzi

Source Tissue

Source Tissue Comment Organism Textmining
amastigote
-
Trypanosoma cruzi
-
epimastigote
-
Trypanosoma cruzi
-
additional information both isozymes appear to be clearly more abundant in the insect stage, although they can be immunodetected in the bloodstream forms Trypanosoma brucei
-
additional information the expression of the mitochondrial isozyme seems to be clearly upregulated in amastigotes, whereas the cytosolic isoform appears to be more abundant in the insect stages of the parasite Trypanosoma cruzi
-
additional information both cytosolic and mitochondrial isozymes are more abundant in the insect stage, although they can be immunodetected in the bloodstream forms Trypanosoma brucei
-
additional information expression of the mitochondrial ME is upregulated in amastigotes, while the cytosolic isoform is more abundant in the insect stages of the parasite Trypanosoma cruzi
-
procyclic form
-
Trypanosoma brucei
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(S)-malate + NADP+
-
Trypanosoma brucei pyruvate + CO2 + NADPH
-
r
(S)-malate + NADP+
-
Trypanosoma cruzi pyruvate + CO2 + NADPH
-
r
(S)-malate + NADP+
-
Trypanosoma cruzi CL Brener pyruvate + CO2 + NADPH
-
r
(S)-malate + NADP+
-
Trypanosoma brucei 427 pyruvate + CO2 + NADPH
-
r
(S)-malate + NADP+
-
Trypanosoma brucei stock 427 pyruvate + CO2 + NADPH
-
r
(S)-malate + NADP+
-
Trypanosoma brucei pyruvate + NADPH + H+ + CO2
-
r
(S)-malate + NADP+
-
Trypanosoma cruzi pyruvate + NADPH + H+ + CO2
-
?
(S)-malate + NADP+ oxaloacetate represents the intermediate resulting from dehydrogenation of malate during the first step of the catalytic cycle of MEs Trypanosoma brucei pyruvate + NADPH + H+ + CO2
-
r
(S)-malate + NADP+ oxaloacetate represents the intermediate resulting from dehydrogenation of malate during the first step of the catalytic cycle of MEs Trypanosoma cruzi pyruvate + NADPH + H+ + CO2
-
r
(S)-malate + NADP+
-
Trypanosoma cruzi CL Brener pyruvate + NADPH + H+ + CO2
-
?
(S)-malate + NADP+ oxaloacetate represents the intermediate resulting from dehydrogenation of malate during the first step of the catalytic cycle of MEs Trypanosoma cruzi CL Brener pyruvate + NADPH + H+ + CO2
-
r
(S)-malate + NADP+
-
Trypanosoma brucei 427 pyruvate + NADPH + H+ + CO2
-
r
(S)-malate + NADP+ oxaloacetate represents the intermediate resulting from dehydrogenation of malate during the first step of the catalytic cycle of MEs Trypanosoma brucei 427 pyruvate + NADPH + H+ + CO2
-
r
(S)-malate + NADP+
-
Trypanosoma brucei stock 427 pyruvate + NADPH + H+ + CO2
-
r
(S)-malate + NADP+ oxaloacetate represents the intermediate resulting from dehydrogenation of malate during the first step of the catalytic cycle of MEs Trypanosoma brucei stock 427 pyruvate + NADPH + H+ + CO2
-
r

Subunits

Subunits Comment Organism
tetramer
-
Trypanosoma brucei
tetramer
-
Trypanosoma cruzi

Synonyms

Synonyms Comment Organism
malic enzyme
-
Trypanosoma brucei
malic enzyme
-
Trypanosoma cruzi

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
8.3
-
pyruvate isozyme ME1, with NADPH and CO2, pH 7.4, temperature not specified in the publication Trypanosoma cruzi
8.3
-
pyruvate pH and temperature not specified in the publication, recombinant isozyme ME1, in presence of 0.5 mM Mn2+ Trypanosoma cruzi
34
-
pyruvate isozyme ME2, with NADPH and CO2, pH 7.4, temperature not specified in the publication Trypanosoma brucei
34
-
pyruvate pH and temperature not specified in the publication, recombinant isozyme ME2, in presence of 0.5 mM Mn2+ Trypanosoma brucei
39
-
pyruvate isozyme ME2, with NADPH and CO2, pH 7.4, temperature not specified in the publication Trypanosoma cruzi
39
-
pyruvate pH and temperature not specified in the publication, recombinant isozyme ME2, in presence of 0.5 mM Mn2+ Trypanosoma cruzi
113
-
pyruvate isozyme ME1, with NADPH and CO2, pH 7.4, temperature not specified in the publication Trypanosoma brucei
113
-
pyruvate pH and temperature not specified in the publication, recombinant isozyme ME1, in presence of 0.5 mM Mn2+ Trypanosoma brucei
242
-
(S)-malate isozyme ME2, pH 7.4, temperature not specified in the publication Trypanosoma brucei
242
-
(S)-malate pH and temperature not specified in the publication, recombinant isozyme ME2, in presence of 0.5 mM Mn2+ Trypanosoma brucei
284
-
NADP+ isozyme ME2, pH 7.4, temperature not specified in the publication Trypanosoma brucei
284
-
NADP+ pH and temperature not specified in the publication, recombinant isozyme ME2, in presence of 0.5 mM Mn2+ Trypanosoma brucei
504
-
NADP+ isozyme ME1, pH 7.4, temperature not specified in the publication Trypanosoma cruzi
504
-
NADP+ pH and temperature not specified in the publication, recombinant isozyme ME1, in presence of 0.5 mM Mn2+ Trypanosoma cruzi
518
-
(S)-malate isozyme ME1, pH 7.4, temperature not specified in the publication Trypanosoma cruzi
518
-
(S)-malate pH and temperature not specified in the publication, recombinant isozyme ME1, in presence of 0.5 mM Mn2+ Trypanosoma cruzi
683
-
(S)-malate isozyme ME2, pH 7.4, temperature not specified in the publication Trypanosoma cruzi
683
-
(S)-malate pH and temperature not specified in the publication, recombinant isozyme ME2, in presence of 0.5 mM Mn2+ Trypanosoma cruzi
691
-
NADP+ isozyme ME2, pH 7.4, temperature not specified in the publication Trypanosoma cruzi
691
-
NADP+ pH and temperature not specified in the publication, recombinant isozyme ME2, in presence of 0.5 mM Mn2+ Trypanosoma cruzi
1132
-
(S)-malate isozyme ME1, pH 7.4, temperature not specified in the publication Trypanosoma brucei
1132
-
(S)-malate pH and temperature not specified in the publication, recombinant isozyme ME1, in presence of 0.5 mM Mn2+ Trypanosoma brucei
1206
-
NADP+ isozyme ME1, pH 7.4, temperature not specified in the publication Trypanosoma brucei
1206
-
NADP+ pH and temperature not specified in the publication, recombinant isozyme ME1, in presence of 0.5 mM Mn2+ Trypanosoma brucei

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.4
-
assay at Trypanosoma brucei
7.4
-
assay at Trypanosoma cruzi

Cofactor

Cofactor Comment Organism Structure
NADP+
-
Trypanosoma brucei
NADP+
-
Trypanosoma cruzi
NADP+ specific for Trypanosoma brucei
NADP+ specific for Trypanosoma cruzi
NADPH
-
Trypanosoma brucei
NADPH
-
Trypanosoma cruzi
NADPH specific for Trypanosoma brucei
NADPH specific for Trypanosoma cruzi

General Information

General Information Comment Organism
physiological function in those environments where glucose is very low or absent, the pathogen depends on NADP-linked dehydrogenases such as the MEs for NADPH production, as in those conditions the pentose phosphate pathway cannot serve as a source of essential reducing power Trypanosoma brucei
physiological function in those environments where glucose is very low or absent, the pathogen depends on NADP-linked dehydrogenases such as the MEs for NADPH production, as in those conditions the pentose phosphate pathway cannot serve as a source of essential reducing power Trypanosoma cruzi
physiological function possibly, in those environments where glucose is very low or absent, the pathogen depends on NADP-linked dehydrogenases such as the MEs for NADPH production, as in those conditions the pentose phosphate pathway cannot serve as a source of essential reducing power Trypanosoma brucei
physiological function possibly, in those environments where glucose is very low or absent, the pathogen depends on NADP-linked dehydrogenases such as the MEs for NADPH production, as in those conditions the pentose phosphate pathway cannot serve as a source of essential reducing power Trypanosoma cruzi

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1
-
pyruvate isozyme ME1, with NADPH and CO2, pH 7.4, temperature not specified in the publication Trypanosoma cruzi
1.1
-
pyruvate pH and temperature not specified in the publication, recombinant isozyme ME1, in presence of 0.5 mM Mn2+ Trypanosoma cruzi
6
-
pyruvate isozyme ME2, with NADPH and CO2, pH 7.4, temperature not specified in the publication Trypanosoma brucei
6
-
pyruvate pH and temperature not specified in the publication, recombinant isozyme ME2, in presence of 0.5 mM Mn2+ Trypanosoma brucei
8
-
pyruvate isozyme ME2, with NADPH and CO2, pH 7.4, temperature not specified in the publication Trypanosoma cruzi
8
-
pyruvate pH and temperature not specified in the publication, recombinant isozyme ME2, in presence of 0.5 mM Mn2+ Trypanosoma cruzi
26
-
pyruvate isozyme ME1, with NADPH and CO2, pH 7.4, temperature not specified in the publication Trypanosoma brucei
26
-
pyruvate pH and temperature not specified in the publication, recombinant isozyme ME1, in presence of 0.5 mM Mn2+ Trypanosoma brucei
297
-
(S)-malate isozyme ME2, pH 7.4, temperature not specified in the publication Trypanosoma cruzi
297
-
(S)-malate pH and temperature not specified in the publication, recombinant isozyme ME2, in presence of 0.5 mM Mn2+ Trypanosoma cruzi
494
-
(S)-malate isozyme ME2, pH 7.4, temperature not specified in the publication Trypanosoma brucei
494
-
(S)-malate pH and temperature not specified in the publication, recombinant isozyme ME2, in presence of 0.5 mM Mn2+ Trypanosoma brucei
3536
-
(S)-malate isozyme ME1, pH 7.4, temperature not specified in the publication Trypanosoma brucei
3536
-
(S)-malate pH and temperature not specified in the publication, recombinant isozyme ME1, in presence of 0.5 mM Mn2+ Trypanosoma brucei
3981
-
(S)-malate isozyme ME1, pH 7.4, temperature not specified in the publication Trypanosoma cruzi
3981
-
(S)-malate pH and temperature not specified in the publication, recombinant isozyme ME1, in presence of 0.5 mM Mn2+ Trypanosoma cruzi
17780
-
NADP+ isozyme ME2, pH 7.4, temperature not specified in the publication Trypanosoma brucei
17780
-
NADP+ pH and temperature not specified in the publication, recombinant isozyme ME2, in presence of 0.5 mM Mn2+ Trypanosoma brucei
18680
-
NADP+ isozyme ME1, pH 7.4, temperature not specified in the publication Trypanosoma cruzi
18680
-
NADP+ pH and temperature not specified in the publication, recombinant isozyme ME1, in presence of 0.5 mM Mn2+ Trypanosoma cruzi
21600
-
NADP+ isozyme ME2, pH 7.4, temperature not specified in the publication Trypanosoma cruzi
21600
-
NADP+ pH and temperature not specified in the publication, recombinant isozyme ME2, in presence of 0.5 mM Mn2+ Trypanosoma cruzi
25170
-
NADP+ isozyme ME1, pH 7.4, temperature not specified in the publication Trypanosoma brucei
25170
-
NADP+ pH and temperature not specified in the publication, recombinant isozyme ME1, in presence of 0.5 mM Mn2+ Trypanosoma brucei