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Literature summary extracted from

  • Serrao, V.; Romanello, L.; Cassago, A.; de Souza, J.; Cheleski, J.; DeMarco, R.; Brandao-Neto, J.; Pereira, H.
    Structure and kinetics assays of recombinant Schistosoma mansoni dihydrofolate reductase (2017), Acta Trop., 170, 190-196 .
    View publication on PubMed

Application

EC Number Application Comment Organism
1.5.1.3 drug development the enzyme is an anti-parasitic drug target, e.g. for malaria or human cancers, because rapidly growing cells require folate to produce thymine Schistosoma mansoni

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.5.1.3 gene Smdhfr, single-copy gene, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) Schistosoma mansoni

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.5.1.3 purified recombinant His-tagged enzyme, mixing of 0.001 ml of 8 mg/ml protein solution with 0.001 ml of well solution containing 100 mM Tris-HCl, pH 8.0, and 2.0 M ammonium sulfate, at 18°C, two weeks, X-ray diffraction structure determination and analysis at 1.95 A resolution, molecular replacement using the modified structure of chicken liver DHFR, PDB ID 8DFR, as search model Schistosoma mansoni

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.5.1.3 aminopterin a folate analogue that is recognized as folate competitor, used as chemotherapeutic agent in cancer and autoimmune diseases Schistosoma mansoni
1.5.1.3 methotrexate a folate analogue that is recognized as folate competitor, used as chemotherapeutic agent in cancer and autoimmune diseases Schistosoma mansoni

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.5.1.3 0.0264
-
 7,8-dihydrofolate pH 7.8, 25°C, recombinant enzyme Schistosoma mansoni
1.5.1.3 0.123
-
 NADP+ pH 7.8, 25°C, recombinant enzyme Schistosoma mansoni

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining

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+ Schistosoma mansoni
-
 5,6,7,8-tetrahydrofolate + NADP+
-
 r

Organism

EC Number Organism UniProt Comment Textmining
1.5.1.3 Schistosoma mansoni G4VJD6
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.5.1.3 recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and dialysis Schistosoma mansoni

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.5.1.3 7,8-dihydrofolate + NADPH + H+
-
 Schistosoma mansoni 5,6,7,8-tetrahydrofolate + NADP+
-
 r
1.5.1.3 additional information substrate binding site analysis, overview Schistosoma mansoni ?
-
 ?

Subunits

EC Number Subunits Comment Organism
1.5.1.3 monomer 1 * 22000, recombinant enzyme, SDS-PAGE, 1 * 21089, sequence calculation Schistosoma mansoni
1.5.1.3 More the DHFR enzyme structure exhibits the canonical DHFR fold Schistosoma mansoni

Synonyms

EC Number Synonyms Comment Organism
1.5.1.3 DHFR
-
 Schistosoma mansoni
1.5.1.3 Smdhfr
-
 Schistosoma mansoni
1.5.1.3 Smp 175230
-
 Schistosoma mansoni

Temperature Optimum [°C]

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

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.5.1.3 35.9
-
 NADP+ pH 7.8, 25°C, recombinant enzyme Schistosoma mansoni
1.5.1.3 130
-
 7,8-dihydrofolate pH 7.8, 25°C, recombinant enzyme Schistosoma mansoni

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.5.1.3 7.8
-
 assay at Schistosoma mansoni

Cofactor

EC Number Cofactor Comment Organism Structure
1.5.1.3 NADP+ binding site analysis, overview Schistosoma mansoni
1.5.1.3 NADPH
-
 Schistosoma mansoni

General Information

EC Number General Information Comment Organism
1.5.1.3 evolution alignment analysis shows several substitutions between Schistosoma mansoni and human DHFR at both the folate- and NADP+-binding sites. The folate-binding site exhibits two differences at three residues (human DHFR numbering): E30D and N64F. The NADP+-binding site exhibits six substitutions at 11 residues: D21G,K54R, K55V, R76S, E77T and S118Y Schistosoma mansoni
1.5.1.3 metabolism dihydrofolate reductase (DHFR) is involved in the thymidylate and is essential for nucleotide metabolism Schistosoma mansoni
1.5.1.3 additional information the DHFR enzyme structure exhibits the canonical DHFR fold Schistosoma mansoni

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.5.1.3 210.56
-
 NADP+ pH 7.8, 25°C, recombinant enzyme Schistosoma mansoni
1.5.1.3 4924.24
-
 7,8-dihydrofolate pH 7.8, 25°C, recombinant enzyme Schistosoma mansoni