Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 3.5.4.9 extracted from

  • Pawelek, P.D.; Allaire, M.; Cygler, M.; MacKenzie, R.E.
    Channeling efficiency in the bifunctional methylenetetrahydrofolate dehydrogenase/cyclohydrolase domain: the effects of site-directed mutagenesis of NADP binding residues (2000), Biochim. Biophys. Acta, 1479, 59-68.
    View publication on PubMed
Search for more literature for 3.5.4.9

Activating Compound

Activating Compound Comment Organism Structure
2',5'-ADP stimulates the cyclohydrolase reverse reaction, 2'-phosphate is involved Homo sapiens

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant enzymes in Escherichia coli JM109 as C-terminally His-tagged enzymes Homo sapiens

Protein Variants

Protein Variants Comment Organism
R173A site-directed mutagenesis, at least 500fold increased Km for NADP+ compared to the wild-type, 163fold increased activity in the forward and reduced activity in the reverse reaction of the cyclohydrolase, unaltered channeling efficiency Homo sapiens
R173E site-directed mutagenesis, no dehydrogenase forward reaction activity, no forward channeling Homo sapiens
R173K site-directed mutagenesis, at least 500fold increased Km for NADP+ compared to the wild-type, 46fold increased activity in the forward and reduced activity in the reverse reaction of the cyclohydrolase, unaltered channeling efficiency Homo sapiens
S197A site-directed mutagenesis, 20fold increased Km for NADP+ compared to the wild-type, reduced activity in the reverse reaction of the cyclohydrolase, no stimulation by 2',5'-ADP, unaltered channeling efficiency Homo sapiens
S197D site-directed mutagenesis, 8.5fold increased activity in the forward reaction of the cyclohydrolase, unaltered channeling efficiency Homo sapiens
S197R site-directed mutagenesis, 10fold increased Km for methenyltetrahydrofolate, reduced activity in the forward and reverse reaction of the cyclohydrolase, unaltered channeling efficiency Homo sapiens
S197T site-directed mutagenesis, slightly increased activity in the forward reaction of the cyclohydrolase, unaltered channeling efficiency Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information the His-tag does not alter the activity and kinetics of the enzymes Homo sapiens
0.016
-
 10-formyltetrahydrofolate reverse reaction, recombinant mutant S197D, substrate 10-formyltetrahydrofolate Homo sapiens
0.018
-
 10-formyltetrahydrofolate reverse reaction, recombinant mutant R173K, substrate 10-formyltetrahydrofolate Homo sapiens
0.019
-
 methenyltetrahydrofolate forward reaction, recombinant mutant S197D Homo sapiens
0.02
-
 10-formyltetrahydrofolate reverse reaction, recombinant mutant S197T, substrate 10-formyltetrahydrofolate Homo sapiens
0.021
-
 methenyltetrahydrofolate forward reaction, recombinant mutant R173A and R173K Homo sapiens
0.024
-
 10-formyltetrahydrofolate reverse reaction, recombinant mutant R173A, substrate 10-formyltetrahydrofolate Homo sapiens
0.025
-
 methenyltetrahydrofolate forward reaction, recombinant mutant S197T Homo sapiens
0.025
-
 methenyltetrahydrofolate forward reaction, recombinant wild-type enzyme Homo sapiens
0.026
-
 methenyltetrahydrofolate forward reaction, recombinant mutant R173E Homo sapiens
0.026
-
 10-formyltetrahydrofolate reverse reaction, recombinant wild-type enzyme, substrate 10-formyltetrahydrofolate Homo sapiens
0.028
-
 10-formyltetrahydrofolate reverse reaction, recombinant mutant S197R, substrate 10-formyltetrahydrofolate Homo sapiens
0.031
-
 10-formyltetrahydrofolate reverse reaction, recombinant mutant R173E, substrate 10-formyltetrahydrofolate Homo sapiens
0.038
-
 methenyltetrahydrofolate forward reaction, recombinant mutant S197A Homo sapiens
0.043
-
 10-formyltetrahydrofolate reverse reaction, recombinant mutant S197A, substrate 10-formyltetrahydrofolate Homo sapiens
0.23
-
 methenyltetrahydrofolate forward reaction, recombinant mutant S197R Homo sapiens

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
 bifunctional methenyltetrahydrofolate dehydrogenase/cyclohydrolase
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli by Ni2+-chelate affinity chromatography Homo sapiens

Reaction

Reaction Comment Organism Reaction ID
5,10-methenyltetrahydrofolate + H2O = 10-formyltetrahydrofolate In eukaryotes, the enzyme occurs as a trifunctional enzyme that also has methylenetetrahydrofolate dehydrogenase (NADP+), EC 1.5.1.5, and formate-tetrahydrofolate ligase, EC 6.3.4.3, activity. In some prokaryotes, it occurs as a bifunctional enzyme, that also has methylenetetrahydrofolate dehydrogenase (NADP+), EC 1.5.1.5, activity or formiminotetrahydrofolate cyclodeaminase, EC 4.3.1.4, activity, residue Arg173 is critical for binding of NADP+ with Ser197 playing a supporting role, both residues interact with the 2'-phosphate of NADP+ Homo sapiens
a

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
 the His-tag does not alter the activity and kinetics of the enzymes, determination of methylenetetrahydrofolate dehydrogenase (NADP+), EC 1.5.1.5, activity, determination of forward channeling in percent Homo sapiens
3.4
-
 forward reaction, recombinant mutant S197R, substrate 5,10-methenyltetrahydrofolate Homo sapiens
6.4
-
 reverse reaction, recombinant mutant S197A, substrate 10-formyltetrahydrofolate, in absence of 2',5'-ADP Homo sapiens
7.2
-
 forward reaction, recombinant wild-type enzyme, substrate 5,10-methenyltetrahydrofolate Homo sapiens
7.3
-
 reverse reaction, recombinant mutant S197R, substrate 10-formyltetrahydrofolate, in absence of 2',5'-ADP Homo sapiens
7.6
-
 reverse reaction, recombinant mutant S197A, substrate 10-formyltetrahydrofolate, in presence of 2',5'-ADP Homo sapiens
8
-
 reverse reaction, recombinant mutant R173A, substrate 10-formyltetrahydrofolate, in presence of 2',5'-ADP Homo sapiens
9
-
 reverse reaction, recombinant mutant R173K, substrate 10-formyltetrahydrofolate, in absence of 2',5'-ADP Homo sapiens
9.2
-
 reverse reaction, recombinant mutant R173E, substrate 10-formyltetrahydrofolate, in presence of 2',5'-ADP Homo sapiens
9.2
-
 reverse reaction, recombinant mutant S197T, substrate 10-formyltetrahydrofolate, in absence of 2',5'-ADP Homo sapiens
9.3
-
 reverse reaction, recombinant mutant S197D, substrate 10-formyltetrahydrofolate, in absence of 2',5'-ADP Homo sapiens
9.6
-
 reverse reaction, recombinant wild-type enzyme, substrate 10-formyltetrahydrofolate, in absence of 2',5'-ADP Homo sapiens
10.8
-
 forward reaction, recombinant mutant S197T, substrate 5,10-methenyltetrahydrofolate Homo sapiens
10.8
-
 reverse reaction, recombinant mutant S197D, substrate 10-formyltetrahydrofolate, in presence of 2',5'-ADP Homo sapiens
12.8
-
 reverse reaction, recombinant mutant R173K, substrate 10-formyltetrahydrofolate, in presence of 2',5'-ADP Homo sapiens
13.5
-
 forward reaction, recombinant mutant S197A, substrate 5,10-methenyltetrahydrofolate Homo sapiens
15.9
-
 reverse reaction, recombinant mutant S197T, substrate 10-formyltetrahydrofolate, in presence of 2',5'-ADP Homo sapiens
22.4
-
 reverse reaction, recombinant wild-type enzyme, substrate 10-formyltetrahydrofolate, in presence of 2',5'-ADP Homo sapiens
22.8
-
 reverse reaction, recombinant mutant S197R, substrate 10-formyltetrahydrofolate, in presence of 2',5'-ADP Homo sapiens
61.6
-
 forward reaction, recombinant mutant S197D, substrate 5,10-methenyltetrahydrofolate Homo sapiens
329
-
 forward reaction, recombinant mutant R173K, substrate 5,10-methenyltetrahydrofolate Homo sapiens
1173
-
 forward reaction, recombinant mutant R173A, substrate 5,10-methenyltetrahydrofolate Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5,10-methenyltetrahydrofolate + H2O 5,10-methenyltetrahydrofolate 5-hydrolase (decyclizing), EC 3.5.4.9, activity of the enzyme Homo sapiens 10-formyltetrahydrofolate
-
 r
5,10-methylenetetrahydrofolate + NADP+ methylenetetrahydrofolate dehydrogenase (NADP+), EC 1.5.1.5, activity of the enzyme Homo sapiens 5,10-methenyltetrahydrofolate + NADPH + H+
-
 r
additional information the enzyme has a bifunctional DC domain that binds NADP+ and one folate molecule within a single catalytic cleft being part of a trifunctional enzyme, substrate channeling with high efficiency independent of NADP+ substrate binding efficiency Homo sapiens ?
-
 ?

Synonyms

Synonyms Comment Organism
methenyltetrahydrofolate dehydrogenase/cyclohydrolase
-
 Homo sapiens
methyleneH4folate cyclohydrolase
-
 Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
107
-
 methenyltetrahydrofolate forward reaction, recombinant mutant S197T Homo sapiens
118
-
 methenyltetrahydrofolate forward reaction, recombinant mutant S197A Homo sapiens
123
-
 methenyltetrahydrofolate forward reaction, recombinant mutant R173E Homo sapiens
128
-
 methenyltetrahydrofolate forward reaction, recombinant mutant R173A Homo sapiens
134
-
 methenyltetrahydrofolate forward reaction, recombinant wild-type enzyme Homo sapiens
141
-
 methenyltetrahydrofolate forward reaction, recombinant mutant R173K Homo sapiens
177
-
 methenyltetrahydrofolate forward reaction, recombinant mutant S197D Homo sapiens
185
-
 methenyltetrahydrofolate forward reaction, recombinant mutant S197R Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.3
-
 assay at Homo sapiens