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Literature summary for 1.3.98.1 extracted from

  • Sariego, I.; Annoura, T.; Nara, T.; Hashimoto, M.; Tsubouchi, A.; Iizumi, K.; Makiuchi, T.; Murata, E.; Kita, K.; Aoki, T.
    Genetic diversity and kinetic properties of Trypanosoma cruzi dihydroorotate dehydrogenase isoforms (2006), Parasitol. Int., 55, 11-16.
    View publication on PubMed

Application

Application Comment Organism
medicine despite their genetic variations, kinetic properties of the three DHODs are conserved, these findings facilitate further exploitation of Trypanosoma cruzi DHOD inhibitors, as chemotherapeutic agents against Chagas' disease Trypanosoma cruzi

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Trypanosoma cruzi
genes subcloned int the EcoRI site of pUC18, PCR products cloned in vector pET100/D-TOPO, recombinant DHOD1 and DHOD2 expressed in Escherichia coli BL21-CodonPlus (DE3)-RP Trypanosoma cruzi

Inhibitors

Inhibitors Comment Organism Structure
Orotate competitively inhibits all three DHOD enzymes to a comparable level Trypanosoma cruzi

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0243
-
dihydroorotate DHOD at 25°C Trypanosoma cruzi
0.0259
-
dihydroorotate native DHOD at 25°C Trypanosoma cruzi
0.03
-
fumarate 37°C, pH 7.0 Trypanosoma cruzi
0.03
-
dihydroorotate 37°C, pH 7.0 Trypanosoma cruzi
0.03
-
dihydroorotate DHOD1 and DHOD2 at 37°C Trypanosoma cruzi
0.03
-
fumarate DHOD1 at 37°C Trypanosoma cruzi
0.035
-
fumarate 37°C, pH 7.0 Trypanosoma cruzi
0.035
-
fumarate DHOD2 at 37°C Trypanosoma cruzi
0.0439
-
fumarate DHOD at 25°C Trypanosoma cruzi
0.0534
-
fumarate native DHOD at 25°C Trypanosoma cruzi
0.067
-
fumarate 37°C, pH 7.0 Trypanosoma cruzi
0.067
-
fumarate DHOD3 at 37°C Trypanosoma cruzi
0.071
-
dihydroorotate 37°C, pH 7.0 Trypanosoma cruzi
0.071
-
dihydroorotate DHOD3 at 37°C Trypanosoma cruzi

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
34100
-
DHOD3, sequence analysis Trypanosoma cruzi
34200
-
DHOD1 and DHOD2, sequence analysis Trypanosoma cruzi
37000
-
recombinant DHOD1, DHOD2 and DHOD3 with an N-terminal His6-tag, affinity chromatography Trypanosoma cruzi

Organism

Organism UniProt Comment Textmining
Trypanosoma cruzi
-
-
-
Trypanosoma cruzi Q4D3W2
-
-
Trypanosoma cruzi Q4DEJ0
-
-
Trypanosoma cruzi Q4DGV2
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant DHOD1, DHOD2 and DHOD3 with an N-terminal His6-tag, by affinity chromatography Trypanosoma cruzi

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
dihydroorotate + fumarate
-
Trypanosoma cruzi orotate + ?
-
?
dihydroorotate + fumarate
-
Trypanosoma cruzi orotate + reduced fumarate
-
?

Synonyms

Synonyms Comment Organism
DHOD1
-
Trypanosoma cruzi
DHOD2
-
Trypanosoma cruzi
DHOD3
-
Trypanosoma cruzi

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
-
Trypanosoma cruzi
7
-
DHOD1, DHOD2 and DHOD3 Trypanosoma cruzi

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.0156
-
Orotate inhibition of DHOD2 at 25°C, in the presence of 1 mM fumarate Trypanosoma cruzi
0.0205
-
Orotate inhibition of DHOD2 at 25°C, in the presence of 1 mM dihydroorotate Trypanosoma cruzi
0.0424
-
Orotate 37°C, pH 7.0, versus dihydroorotate Trypanosoma cruzi
0.0424
-
Orotate inhibition of DHOD3 at 37°C, in the presence of 1 mM dihydroorotate Trypanosoma cruzi
0.049
-
Orotate 37°C, pH 7.0, versus fumarate Trypanosoma cruzi
0.049
-
Orotate inhibition of DHOD3 at 37°C, in the presence of 1 mM fumarate Trypanosoma cruzi
0.0573
-
Orotate 37°C, pH 7.0, versus dihydroorotate Trypanosoma cruzi
0.0573
-
Orotate inhibition of DHOD1 at 37°C, in the presence of 1 mM dihydroorotate Trypanosoma cruzi
0.0622
-
Orotate 37°C, pH 7.0, versus fumarate Trypanosoma cruzi
0.0622
-
Orotate inhibition of DHOD1 at 37°C, in the presence of 1 mM fumarate Trypanosoma cruzi
0.0714
-
Orotate 37°C, pH 7.0, versus dihydroorotate Trypanosoma cruzi
0.0714
-
Orotate inhibition of DHOD2 at 37°C, in the presence of 1 mM dihydroorotate Trypanosoma cruzi
0.0773
-
Orotate 37°C, pH 7.0, versus fumarate Trypanosoma cruzi
0.0773
-
Orotate inhibition of DHOD2 at 37°C, in the presence of 1 mM fumarate Trypanosoma cruzi