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Literature summary 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)

EC Number Cloned (Comment) Organism
1.5.1.3 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
2.1.1.45 expression in Escherichia coli Babesia gibsoni

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.5.1.3 methotrexate
-
Babesia gibsoni
1.5.1.3 pyrimethamine
-
Babesia gibsoni
1.5.1.3 trimethoprim
-
Babesia gibsoni
2.1.1.45 antifolate inhibits activitiy of bifunctional DHFR-TS enzyme Babesia gibsoni

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.5.1.3 0.0016
-
7,8-dihydrofolate 25°C, recombinant GST-tagged enzyme Babesia gibsoni
1.5.1.3 0.0047
-
7,8-dihydrofolate 25°C, recombinant detagged enzyme Babesia gibsoni
1.5.1.3 0.00975
-
NADPH 25°C, recombinant detagged enzyme Babesia gibsoni
1.5.1.3 0.017
-
NADPH 25°C, recombinant GST-tagged enzyme Babesia gibsoni

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
1.5.1.3 cytoplasm
-
Babesia gibsoni 5737
-
2.1.1.45 cytoplasm
-
Babesia gibsoni 5737
-
2.1.1.45 soluble
-
Babesia gibsoni
-
-

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.5.1.3 57000
-
x * 57000, recombinant detagged enzyme, SDS-PAGE Babesia gibsoni
2.1.1.45 58000
-
SDS-PAGE, bifunctional native BgDHFR-TS enzyme from the parasite lysate Babesia gibsoni
2.1.1.45 58800
-
x * 58800, calculated, 57000, SDS-PAGE Babesia gibsoni

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.5.1.3 7,8-dihydrofolate + NADPH + H+ Babesia gibsoni
-
5,6,7,8-tetrahydrofolate + NADP+
-
?
2.1.1.45 5,10-methylenetetrahydrofolate + deoxyuridine monophosphate Babesia gibsoni
-
7,8-dihydrofolate + deoxythymidine monophosphate
-
?

Organism

EC Number Organism UniProt Comment Textmining
1.5.1.3 Babesia gibsoni B5U9U8 gene dhfr-ts
-
2.1.1.45 Babesia gibsoni
-
bifunctional dihydrofolate reductase-thymidylate synthase
-
2.1.1.45 Babesia gibsoni B5U9U8
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.5.1.3 recombinant GST-tagged enzyme from Escherichia coli by glutathione affinity chromatography, the tag is cleaved off Babesia gibsoni
2.1.1.45 glutathione elusion method Babesia gibsoni
2.1.1.45 recombinant protein Babesia gibsoni

Source Tissue

EC Number Source Tissue Comment Organism Textmining
2.1.1.45 merozoite
-
Babesia gibsoni
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.5.1.3 7,8-dihydrofolate + NADPH + H+
-
Babesia gibsoni 5,6,7,8-tetrahydrofolate + NADP+
-
?
1.5.1.3 additional information a bifunctional enzyme with dihydrofolate reductase and thymidylate synthase activities, overview Babesia gibsoni ?
-
?
2.1.1.45 5,10-methylenetetrahydrofolate + deoxyuridine monophosphate
-
Babesia gibsoni 7,8-dihydrofolate + deoxythymidine monophosphate
-
?

Subunits

EC Number Subunits Comment Organism
1.5.1.3 ? x * 57000, recombinant detagged enzyme, SDS-PAGE Babesia gibsoni
2.1.1.45 ? x * 58800, calculated, 57000, SDS-PAGE Babesia gibsoni

Synonyms

EC Number Synonyms Comment Organism
1.5.1.3 DHFR-TS
-
Babesia gibsoni
1.5.1.3 dihydrofolate reductase-thymidylate synthase
-
Babesia gibsoni
2.1.1.45 BgDHFR-TS
-
Babesia gibsoni
2.1.1.45 DHFR-TS
-
Babesia gibsoni
2.1.1.45 dihydrofolate reductase-thymidylate synthase
-
Babesia gibsoni

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.5.1.3 25
-
assay at Babesia gibsoni

Cofactor

EC Number Cofactor Comment Organism Structure
1.5.1.3 NADPH
-
Babesia gibsoni

Ki Value [mM]

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

IC50 Value

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
1.5.1.3 0.0000686
-
25°C, recombinant detagged enzyme Babesia gibsoni methotrexate
1.5.1.3 0.05
-
25°C, recombinant detagged enzyme Babesia gibsoni trimethoprim
1.5.1.3 0.055
-
25°C, recombinant detagged enzyme Babesia gibsoni pyrimethamine