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Literature summary for 3.5.4.9 extracted from

  • Eadsforth, T.C.; Cameron, S.; Hunter, W.N.
    The crystal structure of Leishmania major N5,N10-methylenetetrahydrofolate dehydrogenase/cyclohydrolase and assessment of a potential drug target (2012), Mol. Biochem. Parasitol., 181, 178-185.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.5.4.9

Application

Application Comment Organism
drug development the bifunctional enzyme is a target for design of parasite-specific inhibitors as structure-based drugs Leishmania major

Cloned(Commentary)

Cloned (Comment) Organism
expression of the His-tagged enzyme in Escherichia coli Leishmania major

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant apo-DHCH, hanging drop vapor diffusion, mixing of 0.002 ml 9 mg/ml protein in 50 mM Tris–HCl, 100 mM NaCl, pH 7.5, with 0.002 ml reservoir solution containing 20% PEG 3350 and 0.2 M ammonium acetate, equilibration against 0.07 ml of reservoir solution at 22°C, 3 days, X-ray diffraction structure determination and analysis at 2.7 A resolution, modelling Leishmania major

Inhibitors

Inhibitors Comment Organism Structure
5,6,7,8-tetrahydro-N5,N10-carbonylfolic acid i.e. LY354899 Leishmania major
di-2'-deoxyadenosine 5',5''-P1,P5,alpha,beta-methylene-delta,epsilon-methylene-pentaphosphate-gamma-borano
-
 Leishmania major
di-2'-deoxyadenosine 5',5''-P1,P5,alpha,beta-methylene-gamma,delta-methylene-tetraphosphate
-
 Leishmania major
diadenosine 5',5''-P1,P5,alpha,beta-methylene-delta,epsilon-methylene-pentaphosphate-gamma-borano
-
 Leishmania major
P1,P4-bis(5'-adenosyl)-alpha,beta-gamma,delta-bismethylene-tetraphosphate
-
 Leishmania major

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
65000
-
 recombinant detagged enzyme, gel filtration Leishmania major

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
additional information Leishmania major a bifuncitonal enzyme exhibiting N5,N10-methylenetetrahydrofolate dehydrogenase, EC 1.5.1.5, and N5,N10-methenyltetrahydrofolate cyclohydrolase activities ?
-
 ?
additional information Leishmania major Fridlin a bifuncitonal enzyme exhibiting N5,N10-methylenetetrahydrofolate dehydrogenase, EC 1.5.1.5, and N5,N10-methenyltetrahydrofolate cyclohydrolase activities ?
-
 ?
N5,N10-methenyltetrahydrofolate + H2O Leishmania major
-
 N10-formyltetrahydrofolate
-
 ?
N5,N10-methenyltetrahydrofolate + H2O Leishmania major Fridlin
-
 N10-formyltetrahydrofolate
-
 ?

Organism

Organism UniProt Comment Textmining
Leishmania major Q4Q9F9
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme by nickel affinity chromatography, cleavage of the His-tag by TEV protease, His affinity chromatography, and gel filtration, to over 95% purity Leishmania major

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information a bifuncitonal enzyme exhibiting N5,N10-methylenetetrahydrofolate dehydrogenase, EC 1.5.1.5, and N5,N10-methenyltetrahydrofolate cyclohydrolase activities Leishmania major ?
-
 ?
additional information a bifuncitonal enzyme exhibiting N5,N10-methylenetetrahydrofolate dehydrogenase, EC 1.5.1.5, and N5,N10-methenyltetrahydrofolate cyclohydrolase activities Leishmania major Fridlin ?
-
 ?
N5,N10-methenyltetrahydrofolate + H2O
-
 Leishmania major N10-formyltetrahydrofolate
-
 ?
N5,N10-methenyltetrahydrofolate + H2O
-
 Leishmania major Fridlin N10-formyltetrahydrofolate
-
 ?

Subunits

Subunits Comment Organism
homodimer
-
 Leishmania major

Synonyms

Synonyms Comment Organism
DHCH
-
 Leishmania major
N5,N10-methylenetetrahydrofolate dehydrogenase/cyclohydrolase
-
 Leishmania major
N5,N10-methylenetetrahydrofolate dehydrogenase/N5,N10-methenyltetrahydrofolate cyclohydrolase
-
 Leishmania major

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.009
-
 di-2'-deoxyadenosine 5',5''-P1,P5,alpha,beta-methylene-delta,epsilon-methylene-pentaphosphate-gamma-borano with NPP1, pH 7.3, 37°C Leishmania major
0.013
-
 P1,P4-bis(5'-adenosyl)-alpha,beta-gamma,delta-bismethylene-tetraphosphate with NPP1, pH 7.3, 37°C Leishmania major
0.018
-
 5,6,7,8-tetrahydro-N5,N10-carbonylfolic acid , pH 7.3, 37°C Leishmania major
0.02
-
 diadenosine 5',5''-P1,P5,alpha,beta-methylene-delta,epsilon-methylene-pentaphosphate-gamma-borano with NPP1, pH 7.3, 37°C Leishmania major
0.051
-
 di-2'-deoxyadenosine 5',5''-P1,P5,alpha,beta-methylene-gamma,delta-methylene-tetraphosphate with NPP1, pH 7.3, 37°C Leishmania major

IC50 Value

IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
0.013
-
 with NPP1, pH 7.3, 37°C Leishmania major di-2'-deoxyadenosine 5',5''-P1,P5,alpha,beta-methylene-delta,epsilon-methylene-pentaphosphate-gamma-borano
0.033
-
 with NPP1, pH 7.3, 37°C Leishmania major P1,P4-bis(5'-adenosyl)-alpha,beta-gamma,delta-bismethylene-tetraphosphate
0.05
-
 with NPP1, pH 7.3, 37°C Leishmania major di-2'-deoxyadenosine 5',5''-P1,P5,alpha,beta-methylene-gamma,delta-methylene-tetraphosphate
0.063
-
 with NPP1, pH 7.3, 37°C Leishmania major diadenosine 5',5''-P1,P5,alpha,beta-methylene-delta,epsilon-methylene-pentaphosphate-gamma-borano

General Information

General Information Comment Organism
metabolism three enzyme activities in the protozoan Leishmania major, namely N5,N10-methylenetetrahydrofolate dehydrogenase/N5,N10-methenyltetrahydrofolate cyclohydrolase and N10-formyltetrahydrofolate ligase produce the essential intermediate N10-formyltetrahydrofolate, folate metabolism pathway, overview Leishmania major
additional information active site structure, overview Leishmania major