Activating Compound | Comment | Organism | Structure |
---|---|---|---|
2',5'-ADP | stimulates the cyclohydrolase reverse reaction, 2'-phosphate is involved | Homo sapiens |
Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli JM109 as C-terminally His-tagged enzymes | Homo sapiens |
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 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 | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | - |
bifunctional methenyltetrahydrofolate dehydrogenase/cyclohydrolase | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli by Ni2+-chelate affinity chromatography | Homo sapiens |
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 |
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 | 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 | Comment | Organism |
---|---|---|
methenyltetrahydrofolate dehydrogenase/cyclohydrolase | - |
Homo sapiens |
methyleneH4folate cyclohydrolase | - |
Homo sapiens |
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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.3 | - |
assay at | Homo sapiens |