BRENDA - Enzyme Database
show all sequences of 1.5.1.5

Methylene tetrahydrofolate dehydrogenase/cyclohydrolase and the synthesis of 10-CHO-THF are essential in Leishmania major

Murta, S.M.; Vickers, T.J.; Scott, D.A.; Beverley, S.M.; Mol. Microbiol. 71, 1386-1401 (2009)

Data extracted from this reference:

Application
Application
Commentary
Organism
medicine
Leishmania major is shown to require 10-formyl tetrahydrofolate metabolism, thus this is a possible target for chemotherapy in Leishmania major and potential other protozoan parasites
Leishmania major
Cloned(Commentary)
Cloned (Commentary)
Organism
hexa-His tagged DHCH1 is expressed in Escherichia coli, production of a genetic knock-out; PCR-amplification, expressed in Escherichia coli, overexpression and null-mutant production
Leishmania major
Engineering
Protein Variants
Commentary
Organism
additional information
successful double replacement of the DHCH1 gene is a result of double electroporation with two replacement fragments but an intact gene copy is retained (probably aneuploidy), additional metabolic complementation to bypass the requirement for DHCH1 is not successful either, ectopic addition of DHCH1 or formate-tetrahydrofolate ligase before the second replacement step finally results in a null mutant without chromosomal DHCH1 gene (dhch1-/pXNG4-DHCH1 or dhch1-/pXNG4-FTL) or addition to wild-type in overexpressing transfectants (WT/pXNG4-DHCH1 or WT/pXNG4-FTL)
Leishmania major
Inhibitors
Inhibitors
Commentary
Organism
Structure
(2S)-2-[[4-[(6aR)-3-amino-1,9-dioxo-5,6,6a,7-tetrahydro-4H-imidazol[3,4-f]pteridin-8-yl]benzoyl]amino]pentanedioate
-
Leishmania major
(2S)-2-[[4-[(6aR)-3-amino-1,9-dioxo-5,6,6a,7-tetrahydro-4H-imidazol[3,4-f]pteridin-8-yl]benzoyl]amino]pentanedioic acid
5,10-formyltetrahydrofolate substrate analogue inhibits dehydrogenase activity
Leishmania major
(2S)-2-[[4-[(6aR)-3-amino-1,9-dioxo-5,6,6a,7-tetrahydro-4H-imidazo[3,4-f]pteridin-8-yl]benzoyl]amino]pentanedioic acid
competitive substrate analogue
Leishmania major
2,4-diamino-6-(3,4-dichlorophenoxy)-quinazoline
-
Leishmania major
2,4-diamino-6-benzyl-5-(3-phenylpropyl)-pyrimidine
-
Leishmania major
methotrexate
-
Leishmania major
additional information
no inhibition by pteridine analogues, such as methotrexate, or by hydrophobic diamino-quinazoline, -pyrimidine or -pteridine analogues, such as 2,4-diamino-6-(3,4-dichlorophenoxy)-quinazoline, or 2,4-diamino-6-benzyl-5-(3-phenylpropyl)-pyrimidine, or 2,4-diamino-6,7-diisopropylpteridine
Leishmania major
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.038
-
NADP+
25 mM MOPS pH 7.3, 2.5 mM formaldehyde, 30 mM 2-mercaptoethanol, 27°C, with 250 microM 5,10-methylene-tetrahydrofolate; recombinant DHCH1
Leishmania major
0.12
-
5,10-methylene-tetrahydrofolate
25 mM MOPS pH 7.3, 2.5 mM formaldehyde, 1 mM NADP+, 30 mM 2-mercaptoethanol, 27°C; recombinant DHCH1
Leishmania major
Localization
Localization
Commentary
Organism
GeneOntology No.
Textmining
cytosol
; not in mitochondria as in other organisms
Leishmania major
5829
-
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
5,10-methylene tetrahydrofolate + NADP+
Leishmania major
DH activity
5,10-methenyl tetrahydrofolate + NADPH + H+
-
-
r
5,10-methylene-tetrahydrofolate + NADP+
Leishmania major
-
5,10-methenyl-tetrahydrofolate + NADPH + H+
10-formyl tetrahydrofolate (10-CHO-THF)is the product of subsequently active methenyltetrahydrofolate cyclohydrolase, ATP-dependent formate-tetrahydrofolate ligase catalyzes an alternative one step reaction from tetrahydrofolate + format to 10-formyl tetrahydrofolate
-
?
additional information
Leishmania major
dehydrogenase and cyclohydrogenase activities are contained in the single bifunctional protein DHCH1
?
-
-
-
Organism
Organism
UniProt
Commentary
Textmining
Leishmania major
-
-
-
Purification (Commentary)
Purification (Commentary)
Organism
metal affinity chromatography; recombinant DHCH1 protein is purified by metal-affinity chromatography
Leishmania major
Specific Activity [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
22
-
25 mM MOPS pH 7.3, 2.5 mM formaldehyde, 1 mM NADP+, 30 mM 2-mercaptoethanol, 27°C, 6.3 microM/min/mg activity of the methenyltetrahydrofolate cyclohydrolase; dehydrogenase activity is measured by following the conversion of 5,10-methylenetetrahydrofolate to 5,10-methenyltetrahydrofolate
Leishmania major
Storage Stability
Storage Stability
Organism
10% glycerol required to prevent loss of activity during storage
Leishmania major
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
5,10-methylene tetrahydrofolate + NADP+
DH activity
700281
Leishmania major
5,10-methenyl tetrahydrofolate + NADPH + H+
-
-
-
r
5,10-methylene-tetrahydrofolate + NADP+
-
700281
Leishmania major
5,10-methenyl-tetrahydrofolate + NADPH + H+
10-formyl tetrahydrofolate (10-CHO-THF)is the product of subsequently active methenyltetrahydrofolate cyclohydrolase, ATP-dependent formate-tetrahydrofolate ligase catalyzes an alternative one step reaction from tetrahydrofolate + format to 10-formyl tetrahydrofolate
-
-
?
additional information
dehydrogenase and cyclohydrogenase activities are contained in the single bifunctional protein DHCH1
700281
Leishmania major
?
-
-
-
-
Synonyms
Synonyms
Commentary
Organism
5,10-mehtylene tetrahydrofolate dehydrogenase
-
Leishmania major
5,10-methenyltetrahydrofolate cyclohydrolase/5,10-methylene tetrahydrofolate dehydrogenase
-
Leishmania major
DHCH1
bifuntional enzyme with 5,10-mehtylene tetrahydrofolate dehydrogenase and methenyltetrahydrofolate cyclohydrolase activity in Leishmania major
Leishmania major
methylene tetrahydrofolate dehydrogenase/cyclohydrolase
-
Leishmania major
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
27
-
assay at
Leishmania major
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
10
-
NADP+
25 mM MOPS pH 7.3, 2.5 mM formaldehyde, 30 mM 2-mercaptoethanol, 27°C; recombinant DHCH1, 1 mM NADP+
Leishmania major
14
-
5,10-methylene-tetrahydrofolate
25 mM MOPS pH 7.3, 2.5 mM formaldehyde, 1 mM NADP+, 30 mM 2-mercaptoethanol, 27°C
Leishmania major
14
-
5,10-methylene tetrahydrofolate
recombinant DHCH1, 0.25 mM substrate
Leishmania major
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.3
-
assay at
Leishmania major
Cofactor
Cofactor
Commentary
Organism
Structure
NADP+
; no dependence on NAD+ in presence of K3PO4 and MgCl2 (required for human mitochondrial NAD+ dependent activity of bifunctional dehydrogenase cyclase)
Leishmania major
Ki Value [mM]
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
0.000105
-
(2S)-2-[[4-[(6aR)-3-amino-1,9-dioxo-5,6,6a,7-tetrahydro-4H-imidazo[3,4-f]pteridin-8-yl]benzoyl]amino]pentanedioic acid
pH 7.3, 27°C, Leishmania wild-type growth is inhibited with an EC50 of 1.1 microM, DHCH1 or formate-tetrahydrofolate ligase overexpressing (FTL) Leishmania transfectants show 4-5fold or 3fold resistance, respectively
Leishmania major
0.000105
-
(2S)-2-[[4-[(6aR)-3-amino-1,9-dioxo-5,6,6a,7-tetrahydro-4H-imidazol[3,4-f]pteridin-8-yl]benzoyl]amino]pentanedioate
-
Leishmania major
IC50 Value
IC50 Value
IC50 Value Maximum
Commentary
Organism
Inhibitor
Structure
0.000036
-
wild type enzyme
Leishmania major
methotrexate
0.000041
-
dhch1-/pXNG4-DHCH1 mutant, FV1 line
Leishmania major
methotrexate
0.0004
-
dhch1-/pXNG4-DHCH1 mutant, FV1 line
Leishmania major
2,4-diamino-6-benzyl-5-(3-phenylpropyl)-pyrimidine
0.00044
-
wild type enzyme
Leishmania major
2,4-diamino-6-benzyl-5-(3-phenylpropyl)-pyrimidine
0.00068
-
wild type enzyme
Leishmania major
2,4-diamino-6-(3,4-dichlorophenoxy)-quinazoline
0.00076
-
dhch1-/pXNG4-DHCH1 mutant, FV1 line
Leishmania major
2,4-diamino-6-(3,4-dichlorophenoxy)-quinazoline
0.0011
-
50% growth inhibition of wild-type cell line
Leishmania major
(2S)-2-[[4-[(6aR)-3-amino-1,9-dioxo-5,6,6a,7-tetrahydro-4H-imidazo[3,4-f]pteridin-8-yl]benzoyl]amino]pentanedioic acid
0.0011
-
wild type enzyme
Leishmania major
(2S)-2-[[4-[(6aR)-3-amino-1,9-dioxo-5,6,6a,7-tetrahydro-4H-imidazol[3,4-f]pteridin-8-yl]benzoyl]amino]pentanedioic acid
0.0033
-
50% growth inhibition of null mutant with plasmid expressing formate-tetrahydrofolate ligase
Leishmania major
(2S)-2-[[4-[(6aR)-3-amino-1,9-dioxo-5,6,6a,7-tetrahydro-4H-imidazo[3,4-f]pteridin-8-yl]benzoyl]amino]pentanedioic acid
0.0037
-
50% growth inhibition of wild-type with plasmid expressing formate-tetrahydrofolate ligase
Leishmania major
(2S)-2-[[4-[(6aR)-3-amino-1,9-dioxo-5,6,6a,7-tetrahydro-4H-imidazo[3,4-f]pteridin-8-yl]benzoyl]amino]pentanedioic acid
0.0045
-
50% growth inhibition of null mutant with plasmid expressing the enzyme
Leishmania major
(2S)-2-[[4-[(6aR)-3-amino-1,9-dioxo-5,6,6a,7-tetrahydro-4H-imidazo[3,4-f]pteridin-8-yl]benzoyl]amino]pentanedioic acid
0.0045
-
dhch1-/pXNG4-DHCH1 mutant, FV1 line
Leishmania major
(2S)-2-[[4-[(6aR)-3-amino-1,9-dioxo-5,6,6a,7-tetrahydro-4H-imidazol[3,4-f]pteridin-8-yl]benzoyl]amino]pentanedioic acid
0.0051
-
50% growth inhibition of wild-type with plasmid expressing the enzyme (overexpressing it as a result)
Leishmania major
(2S)-2-[[4-[(6aR)-3-amino-1,9-dioxo-5,6,6a,7-tetrahydro-4H-imidazo[3,4-f]pteridin-8-yl]benzoyl]amino]pentanedioic acid
0.0051
-
WT/pXNG4-DHCH mutant, FV1 line
Leishmania major
(2S)-2-[[4-[(6aR)-3-amino-1,9-dioxo-5,6,6a,7-tetrahydro-4H-imidazol[3,4-f]pteridin-8-yl]benzoyl]amino]pentanedioic acid
Application (protein specific)
Application
Commentary
Organism
medicine
Leishmania major is shown to require 10-formyl tetrahydrofolate metabolism, thus this is a possible target for chemotherapy in Leishmania major and potential other protozoan parasites
Leishmania major
Cloned(Commentary) (protein specific)
Commentary
Organism
hexa-His tagged DHCH1 is expressed in Escherichia coli, production of a genetic knock-out; PCR-amplification, expressed in Escherichia coli, overexpression and null-mutant production
Leishmania major
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NADP+
; no dependence on NAD+ in presence of K3PO4 and MgCl2 (required for human mitochondrial NAD+ dependent activity of bifunctional dehydrogenase cyclase)
Leishmania major
Engineering (protein specific)
Protein Variants
Commentary
Organism
additional information
successful double replacement of the DHCH1 gene is a result of double electroporation with two replacement fragments but an intact gene copy is retained (probably aneuploidy), additional metabolic complementation to bypass the requirement for DHCH1 is not successful either, ectopic addition of DHCH1 or formate-tetrahydrofolate ligase before the second replacement step finally results in a null mutant without chromosomal DHCH1 gene (dhch1-/pXNG4-DHCH1 or dhch1-/pXNG4-FTL) or addition to wild-type in overexpressing transfectants (WT/pXNG4-DHCH1 or WT/pXNG4-FTL)
Leishmania major
IC50 Value (protein specific)
IC50 Value
IC50 Value Maximum
Commentary
Organism
Inhibitor
Structure
0.000036
-
wild type enzyme
Leishmania major
methotrexate
0.000041
-
dhch1-/pXNG4-DHCH1 mutant, FV1 line
Leishmania major
methotrexate
0.0004
-
dhch1-/pXNG4-DHCH1 mutant, FV1 line
Leishmania major
2,4-diamino-6-benzyl-5-(3-phenylpropyl)-pyrimidine
0.00044
-
wild type enzyme
Leishmania major
2,4-diamino-6-benzyl-5-(3-phenylpropyl)-pyrimidine
0.00068
-
wild type enzyme
Leishmania major
2,4-diamino-6-(3,4-dichlorophenoxy)-quinazoline
0.00076
-
dhch1-/pXNG4-DHCH1 mutant, FV1 line
Leishmania major
2,4-diamino-6-(3,4-dichlorophenoxy)-quinazoline
0.0011
-
50% growth inhibition of wild-type cell line
Leishmania major
(2S)-2-[[4-[(6aR)-3-amino-1,9-dioxo-5,6,6a,7-tetrahydro-4H-imidazo[3,4-f]pteridin-8-yl]benzoyl]amino]pentanedioic acid
0.0011
-
wild type enzyme
Leishmania major
(2S)-2-[[4-[(6aR)-3-amino-1,9-dioxo-5,6,6a,7-tetrahydro-4H-imidazol[3,4-f]pteridin-8-yl]benzoyl]amino]pentanedioic acid
0.0033
-
50% growth inhibition of null mutant with plasmid expressing formate-tetrahydrofolate ligase
Leishmania major
(2S)-2-[[4-[(6aR)-3-amino-1,9-dioxo-5,6,6a,7-tetrahydro-4H-imidazo[3,4-f]pteridin-8-yl]benzoyl]amino]pentanedioic acid
0.0037
-
50% growth inhibition of wild-type with plasmid expressing formate-tetrahydrofolate ligase
Leishmania major
(2S)-2-[[4-[(6aR)-3-amino-1,9-dioxo-5,6,6a,7-tetrahydro-4H-imidazo[3,4-f]pteridin-8-yl]benzoyl]amino]pentanedioic acid
0.0045
-
50% growth inhibition of null mutant with plasmid expressing the enzyme
Leishmania major
(2S)-2-[[4-[(6aR)-3-amino-1,9-dioxo-5,6,6a,7-tetrahydro-4H-imidazo[3,4-f]pteridin-8-yl]benzoyl]amino]pentanedioic acid
0.0045
-
dhch1-/pXNG4-DHCH1 mutant, FV1 line
Leishmania major
(2S)-2-[[4-[(6aR)-3-amino-1,9-dioxo-5,6,6a,7-tetrahydro-4H-imidazol[3,4-f]pteridin-8-yl]benzoyl]amino]pentanedioic acid
0.0051
-
50% growth inhibition of wild-type with plasmid expressing the enzyme (overexpressing it as a result)
Leishmania major
(2S)-2-[[4-[(6aR)-3-amino-1,9-dioxo-5,6,6a,7-tetrahydro-4H-imidazo[3,4-f]pteridin-8-yl]benzoyl]amino]pentanedioic acid
0.0051
-
WT/pXNG4-DHCH mutant, FV1 line
Leishmania major
(2S)-2-[[4-[(6aR)-3-amino-1,9-dioxo-5,6,6a,7-tetrahydro-4H-imidazol[3,4-f]pteridin-8-yl]benzoyl]amino]pentanedioic acid
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
(2S)-2-[[4-[(6aR)-3-amino-1,9-dioxo-5,6,6a,7-tetrahydro-4H-imidazol[3,4-f]pteridin-8-yl]benzoyl]amino]pentanedioate
-
Leishmania major
(2S)-2-[[4-[(6aR)-3-amino-1,9-dioxo-5,6,6a,7-tetrahydro-4H-imidazol[3,4-f]pteridin-8-yl]benzoyl]amino]pentanedioic acid
5,10-formyltetrahydrofolate substrate analogue inhibits dehydrogenase activity
Leishmania major
(2S)-2-[[4-[(6aR)-3-amino-1,9-dioxo-5,6,6a,7-tetrahydro-4H-imidazo[3,4-f]pteridin-8-yl]benzoyl]amino]pentanedioic acid
competitive substrate analogue
Leishmania major
2,4-diamino-6-(3,4-dichlorophenoxy)-quinazoline
-
Leishmania major
2,4-diamino-6-benzyl-5-(3-phenylpropyl)-pyrimidine
-
Leishmania major
methotrexate
-
Leishmania major
additional information
no inhibition by pteridine analogues, such as methotrexate, or by hydrophobic diamino-quinazoline, -pyrimidine or -pteridine analogues, such as 2,4-diamino-6-(3,4-dichlorophenoxy)-quinazoline, or 2,4-diamino-6-benzyl-5-(3-phenylpropyl)-pyrimidine, or 2,4-diamino-6,7-diisopropylpteridine
Leishmania major
Ki Value [mM] (protein specific)
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
0.000105
-
(2S)-2-[[4-[(6aR)-3-amino-1,9-dioxo-5,6,6a,7-tetrahydro-4H-imidazo[3,4-f]pteridin-8-yl]benzoyl]amino]pentanedioic acid
pH 7.3, 27°C, Leishmania wild-type growth is inhibited with an EC50 of 1.1 microM, DHCH1 or formate-tetrahydrofolate ligase overexpressing (FTL) Leishmania transfectants show 4-5fold or 3fold resistance, respectively
Leishmania major
0.000105
-
(2S)-2-[[4-[(6aR)-3-amino-1,9-dioxo-5,6,6a,7-tetrahydro-4H-imidazol[3,4-f]pteridin-8-yl]benzoyl]amino]pentanedioate
-
Leishmania major
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.038
-
NADP+
25 mM MOPS pH 7.3, 2.5 mM formaldehyde, 30 mM 2-mercaptoethanol, 27°C, with 250 microM 5,10-methylene-tetrahydrofolate; recombinant DHCH1
Leishmania major
0.12
-
5,10-methylene-tetrahydrofolate
25 mM MOPS pH 7.3, 2.5 mM formaldehyde, 1 mM NADP+, 30 mM 2-mercaptoethanol, 27°C; recombinant DHCH1
Leishmania major
Localization (protein specific)
Localization
Commentary
Organism
GeneOntology No.
Textmining
cytosol
; not in mitochondria as in other organisms
Leishmania major
5829
-
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
5,10-methylene tetrahydrofolate + NADP+
Leishmania major
DH activity
5,10-methenyl tetrahydrofolate + NADPH + H+
-
-
r
5,10-methylene-tetrahydrofolate + NADP+
Leishmania major
-
5,10-methenyl-tetrahydrofolate + NADPH + H+
10-formyl tetrahydrofolate (10-CHO-THF)is the product of subsequently active methenyltetrahydrofolate cyclohydrolase, ATP-dependent formate-tetrahydrofolate ligase catalyzes an alternative one step reaction from tetrahydrofolate + format to 10-formyl tetrahydrofolate
-
?
additional information
Leishmania major
dehydrogenase and cyclohydrogenase activities are contained in the single bifunctional protein DHCH1
?
-
-
-
Purification (Commentary) (protein specific)
Commentary
Organism
metal affinity chromatography; recombinant DHCH1 protein is purified by metal-affinity chromatography
Leishmania major
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
22
-
25 mM MOPS pH 7.3, 2.5 mM formaldehyde, 1 mM NADP+, 30 mM 2-mercaptoethanol, 27°C, 6.3 microM/min/mg activity of the methenyltetrahydrofolate cyclohydrolase; dehydrogenase activity is measured by following the conversion of 5,10-methylenetetrahydrofolate to 5,10-methenyltetrahydrofolate
Leishmania major
Storage Stability (protein specific)
Storage Stability
Organism
10% glycerol required to prevent loss of activity during storage
Leishmania major
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
5,10-methylene tetrahydrofolate + NADP+
DH activity
700281
Leishmania major
5,10-methenyl tetrahydrofolate + NADPH + H+
-
-
-
r
5,10-methylene-tetrahydrofolate + NADP+
-
700281
Leishmania major
5,10-methenyl-tetrahydrofolate + NADPH + H+
10-formyl tetrahydrofolate (10-CHO-THF)is the product of subsequently active methenyltetrahydrofolate cyclohydrolase, ATP-dependent formate-tetrahydrofolate ligase catalyzes an alternative one step reaction from tetrahydrofolate + format to 10-formyl tetrahydrofolate
-
-
?
additional information
dehydrogenase and cyclohydrogenase activities are contained in the single bifunctional protein DHCH1
700281
Leishmania major
?
-
-
-
-
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
27
-
assay at
Leishmania major
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
10
-
NADP+
25 mM MOPS pH 7.3, 2.5 mM formaldehyde, 30 mM 2-mercaptoethanol, 27°C; recombinant DHCH1, 1 mM NADP+
Leishmania major
14
-
5,10-methylene-tetrahydrofolate
25 mM MOPS pH 7.3, 2.5 mM formaldehyde, 1 mM NADP+, 30 mM 2-mercaptoethanol, 27°C
Leishmania major
14
-
5,10-methylene tetrahydrofolate
recombinant DHCH1, 0.25 mM substrate
Leishmania major
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.3
-
assay at
Leishmania major
General Information
General Information
Commentary
Organism
metabolism
product of the bifuncional enzyme, 10-formyl tetrahydrofolate, is not essential for de novo purine synthesis in Leishmania and other parasitic protozoans contrary to other organisms, it is utilized for methionyl-tRNA formulation in mitochondria, the bifunctional enzyme product is essential for Leishmania major and may even reveal novel pathways
Leishmania major
General Information (protein specific)
General Information
Commentary
Organism
metabolism
product of the bifuncional enzyme, 10-formyl tetrahydrofolate, is not essential for de novo purine synthesis in Leishmania and other parasitic protozoans contrary to other organisms, it is utilized for methionyl-tRNA formulation in mitochondria, the bifunctional enzyme product is essential for Leishmania major and may even reveal novel pathways
Leishmania major
KCat/KM [mM/s]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
120
-
5,10-methylene-tetrahydrofolate
25 mM MOPS pH 7.3, 2.5 mM formaldehyde, 1 mM NADP+, 30 mM 2-mercaptoethanol, 27°C
Leishmania major
260
-
NADP+
25 mM MOPS pH 7.3, 2.5 mM formaldehyde, 30 mM 2-mercaptoethanol, 27°C, with 250 microM 5,10-methylene-tetrahydrofolate
Leishmania major
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
120
-
5,10-methylene-tetrahydrofolate
25 mM MOPS pH 7.3, 2.5 mM formaldehyde, 1 mM NADP+, 30 mM 2-mercaptoethanol, 27°C
Leishmania major
260
-
NADP+
25 mM MOPS pH 7.3, 2.5 mM formaldehyde, 30 mM 2-mercaptoethanol, 27°C, with 250 microM 5,10-methylene-tetrahydrofolate
Leishmania major
Other publictions for EC 1.5.1.5
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Synonyms
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
743250
Aluri
Physiological role of FolD (m ...
Clostridium perfringens
Microbiology
162
145-155
2016
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720406
Eadsforth
The crystal structure of Leish ...
Leishmania major, Leishmania major Fridlin
Mol. Biochem. Parasitol.
181
178-185
2012
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718743
Lee
Crystal structure of bifunctio ...
Thermoplasma acidophilum
Biochem. Biophys. Res. Commun.
406
459-463
2011
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725456
Bolusani
Mammalian MTHFD2L encodes a mi ...
Homo sapiens, Rattus norvegicus
J. Biol. Chem.
286
5166-5174
2011
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12
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712807
Bi
Association analysis of CbetaS ...
Homo sapiens
J. Neural Transm.
117
499-503
2010
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698187
Christensen
The MTHFD1 p.Arg653Gln variant ...
Homo sapiens
Hum. Mutat.
30
212-220
2009
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700281
Murta
Methylene tetrahydrofolate deh ...
Leishmania major
Mol. Microbiol.
71
1386-1401
2009
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1
1
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1
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7
2
1
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7
2
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1
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1
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1
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2
2
688323
Kim
Crystallization and preliminar ...
Thermoplasma acidophilum
J. Microbiol. Biotechnol.
18
283-286
2008
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1
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1
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701402
Christensen
Mitochondrial methylenetetrahy ...
Homo sapiens, Saccharomyces cerevisiae
Vitam. Horm.
79
393-410
2008
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2
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3
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1
1
4
3
1
7
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2
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2
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7
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11
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3
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3
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3
1
7
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2
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7
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1
1
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654937
Casaite
-
Elucidation of enzymatic funct ...
Arthrobacter globiformis
Biologia (Bratisl. )
2
1-4
2004
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1
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392334
Goenrich
Purification and characterizat ...
Hyphomicrobium zavarzinii, Hyphomicrobium zavarzinii ZV580, no activity in Methylobacterium extorquens
Arch. Microbiol.
177
299-303
2002
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2
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2
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4
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1
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392333
Pawelek
Methenyltetrahydrofolate cyclo ...
Homo sapiens, Photobacterium phosphoreum
Biochemistry
37
1109-1115
1998
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7
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1
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2
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7
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4
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209738
Pawelek
Methylenetetrahydrofolate dehy ...
Photobacterium phosphoreum
Biochim. Biophys. Acta
1296
47-54
1996
1
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2
4
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209737
Kirk
-
A 5,10-methylenetetrahydrofola ...
Pisum sativum
Phytochemistry
39
1309-1317
1995
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-
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4
3
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2
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1
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3
3
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2
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392331
Pelletier J.N.; MacKenzie
Binding and interconversion of ...
Homo sapiens
Biochemistry
34
12673-12680
1995
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1
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1518
Cossins
Enzymes for synthesis of 10-fo ...
Pisum sativum
Adv. Exp. Med. Biol.
338
707-710
1993
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2
1
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1
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1
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1
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392079
Mackenzie
Methylenetetrahydrofolate dehy ...
Homo sapiens, Mus musculus
Adv. Enzyme Regul.
27
31-39
1988
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5
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392315
Hum
Primary structure of a human t ...
Homo sapiens
J. Biol. Chem.
263
15946-15950
1988
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392323
Gardam
The NADP-dependent trifunction ...
Mus musculus
Biochem. Cell Biol.
66
66-70
1988
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1
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1532
Villar
C1-Tetrahydrofolate synthase f ...
Oryctolagus cuniculus
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260
2245-2252
1985
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392321
Ross
Studies on the polyglutamate s ...
Sus scrofa
Biochemistry
23
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Paukert
Formyl-methenyl-methylenetetra ...
Ovis aries, Saccharomyces cerevisiae, Sus scrofa
Methods Enzymol.
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1980
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2
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3
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3
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1
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3
1
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392309
Ljungdahl
Methylenetetrahydrofolate dehy ...
Moorella thermoacetica
Methods Enzymol.
66
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1980
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1
1
1
1
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392317
MacKenzie
Methylenetetrahydrofolate dehy ...
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Methods Enzymol.
66
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1
1
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1
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209726
Cohen
Methylenetetrahydrofolate dehy ...
Sus scrofa
Biochim. Biophys. Acta
522
311-317
1978
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392307
Dev
A complex of N5,N10-methylenet ...
Escherichia coli, Escherichia coli B / ATCC 11303
J. Biol. Chem.
253
4245-4253
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Tan
Methylenetetrahydrofolate dehy ...
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2
1
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392314
Paukert
Formyl-methenyl-methylenetetra ...
Saccharomyces cerevisiae
Biochem. Biophys. Res. Commun.
77
147-154
1977
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1
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392324
Paukert
Formyl-methyl-methylenetetrahy ...
Ovis aries
J. Biol. Chem.
251
5104-5111
1976
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2
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3
1
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1
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1
1
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1
1
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392308
Harvey
Regulation in the folate pathw ...
Escherichia coli, Escherichia coli B / ATCC 11303
Adv. Enzyme Regul.
13
99-124
1975
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3
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2
-
177
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2
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2
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392311
O'Brien
Purification and characterizat ...
Moorella thermoacetica
J. Biol. Chem.
248
403-408
1973
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1
2
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1
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2
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1
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1
2
1
1
1
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1
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392319
MacKenzie
Co-purification of three folat ...
Sus scrofa
Biochem. Biophys. Res. Commun.
53
1088-1095
1973
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1
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1
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392328
Yoshida
Comparative study on major pat ...
Bos taurus, Elaphe quadrivirgata, Gallus gallus bankiva, Ovis aries
J. Biochem.
72
1503-1516
1972
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8
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4
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4
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8
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4
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8
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4
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4
8
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4
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392326
Cossins
-
Plant N5,N10-methylenetetrahyd ...
Pisum sativum
Phytochemistry
9
1463-1471
1970
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1
3
3
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1
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3
1
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1
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1
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3
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3
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1
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3
1
1
2
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1
1
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392306
Albrecht
Methylenetetrahydrofolate dehy ...
Enterococcus faecium
J. Bacteriol.
95
1779-1789
1968
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1
5
2
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1
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1
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1
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1
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5
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1
1
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1
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392310
Uyeda
Enzymes of clostridial purine ...
Clostridium cylindrosporum
J. Biol. Chem.
242
4378-4385
1967
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1
1
4
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1
1
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1
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1
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1
1
3
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1
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4
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1
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1
1
3
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1
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392312
Dalal
Purification of 5,10-methylene ...
Salmonella enterica subsp. enterica serovar Typhimurium, Salmonella enterica subsp. enterica serovar Typhimurium LT2
J. Biol. Chem.
242
3636-3640
1967
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1
3
2
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2
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44
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1
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2
1
2
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1
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1
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1
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2
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1
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2
1
2
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1
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392316
Yeh
Purification and properties of ...
Bos taurus
Biochim. Biophys. Acta
105
279-291
1965
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1
6
3
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1
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1
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1
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1
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1
3
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1
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1
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1
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1
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6
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3
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1
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1
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1
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1
3
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1
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1
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392313
Ramasastri
5,10-Methylenetetrahydrofolic ...
Saccharomyces cerevisiae
J. Biol. Chem.
237
1982-1988
1962
5
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1
5
2
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1
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1
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1
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1
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1
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1
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1
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1
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1
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5
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1
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1
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5
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2
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1
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1
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1
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1
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