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Literature summary for 1.5.1.3 extracted from

  • Aboge, G.O.; Jia, H.; Terkawi, M.A.; Goo, Y.K.; Nishikawa, Y.; Sunaga, F.; Namikawa, K.; Tsuji, N.; Igarashi, I.; Suzuki, H.; Fujisaki, K.; Xuan, X.
    Cloning, expression and characterization of Babesia gibsoni dihydrofolate reductase-thymidylate synthase: inhibitory effect of antifolates on its catalytic activity and the parasite proliferation (2008), Antimicrob. Agents Chemother., 52, 4072-4080.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
gene dhfr-ts, DNA and amino acid sequence determination and analysis, a 1,548-bp open reading frame encoding a 58.8-kDa translated peptide containing DHFR and TS domains linked together in a single polypeptide chain, expression of the GST-tagged enzyme in Escherichia coli strain BL21(DE3) as soluble protein, expression as GFP-tagged enzyme in Babesia gibsoni cytoplasm Babesia gibsoni

Inhibitors

Inhibitors Comment Organism Structure
methotrexate
-
Babesia gibsoni
pyrimethamine
-
Babesia gibsoni
trimethoprim
-
Babesia gibsoni

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0016
-
7,8-dihydrofolate 25°C, recombinant GST-tagged enzyme Babesia gibsoni
0.0047
-
7,8-dihydrofolate 25°C, recombinant detagged enzyme Babesia gibsoni
0.00975
-
NADPH 25°C, recombinant detagged enzyme Babesia gibsoni
0.017
-
NADPH 25°C, recombinant GST-tagged enzyme Babesia gibsoni

Localization

Localization Comment Organism GeneOntology No. Textmining
cytoplasm
-
Babesia gibsoni 5737
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
57000
-
x * 57000, recombinant detagged enzyme, SDS-PAGE Babesia gibsoni

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
7,8-dihydrofolate + NADPH + H+ Babesia gibsoni
-
5,6,7,8-tetrahydrofolate + NADP+
-
?

Organism

Organism UniProt Comment Textmining
Babesia gibsoni B5U9U8 gene dhfr-ts
-

Purification (Commentary)

Purification (Comment) Organism
recombinant GST-tagged enzyme from Escherichia coli by glutathione affinity chromatography, the tag is cleaved off Babesia gibsoni

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
7,8-dihydrofolate + NADPH + H+
-
Babesia gibsoni 5,6,7,8-tetrahydrofolate + NADP+
-
?
additional information a bifunctional enzyme with dihydrofolate reductase and thymidylate synthase activities, overview Babesia gibsoni ?
-
?

Subunits

Subunits Comment Organism
? x * 57000, recombinant detagged enzyme, SDS-PAGE Babesia gibsoni

Synonyms

Synonyms Comment Organism
DHFR-TS
-
Babesia gibsoni
dihydrofolate reductase-thymidylate synthase
-
Babesia gibsoni

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Babesia gibsoni

Cofactor

Cofactor Comment Organism Structure
NADPH
-
Babesia gibsoni

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
additional information
-
additional information antifolate inhibition kinetics, overview Babesia gibsoni

IC50 Value

IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
0.0000686
-
25°C, recombinant detagged enzyme Babesia gibsoni methotrexate
0.05
-
25°C, recombinant detagged enzyme Babesia gibsoni trimethoprim
0.055
-
25°C, recombinant detagged enzyme Babesia gibsoni pyrimethamine