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Literature summary for 1.5.1.15 extracted from
- Monomeric NADH-oxidizing methylenetetrahydrofolate reductases from Mycobacterium smegmatis lack flavin coenzyme (2020), J. Bacteriol., 202, e00709-19 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene MSMEG_6596, DNA and amino acid sequence determination and analysis, sequence comparisons | Mycolicibacterium smegmatis |
| gene MSMEG_6649, DNA and amino acid sequence determination and analysis, sequence comparisons | Mycolicibacterium smegmatis |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| additional information | construction of the Mycobacterium smegmatis DELTAmsmeg_6596 strain. The mutant is partially auxotrophic for methionine and grows only poorly without methionine or without being complemented with a functional copy of MTHFR1 or MTHFR2. Furthermore, the DELTAmsmeg_6596 strain is more sensitive to folate pathway inhibitors (sulfachloropyridazine, p-aminosalicylic acid, sulfamethoxazole, and trimethoprim) | Mycolicibacterium smegmatis |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0325 | - |
NADH | pH 7.6, 22°C | Mycolicibacterium smegmatis |  |
| 0.0632 | - |
5,10-methenyltetrahydrofolate | pH 7.6, 22°C | Mycolicibacterium smegmatis | |
| 0.108 | - |
NADH | pH 7.6, 22°C | Mycolicibacterium smegmatis | |
| 0.145 | - |
5,10-methenyltetrahydrofolate | pH 7.6, 22°C | Mycolicibacterium smegmatis | |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 5,10-methenyltetrahydrofolate + NADH + H+ | Mycolicibacterium smegmatis | - |
5,10-methylenetetrahydrofolate + NAD+ | - |
ir | |
| 5,10-methenyltetrahydrofolate + NADH + H+ | Mycolicibacterium smegmatis ATCC 700084 | - |
5,10-methylenetetrahydrofolate + NAD+ | - |
ir | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Mycolicibacterium smegmatis | A0R6M0 | - |
- |
| Mycolicibacterium smegmatis | A0R6S0 | - |
- |
| Mycolicibacterium smegmatis ATCC 700084 | A0R6M0 | - |
- |
| Mycolicibacterium smegmatis ATCC 700084 | A0R6S0 | - |
- |
| no activity by Mycobacterium smegmatis MSMEG_6664 | - |
- |
- |
| no activity by Mycobacterium smegmatis MSMEI_6484 | - |
- |
- |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 5,10-methenyltetrahydrofolate + NADH + H+ | - |
Mycolicibacterium smegmatis | 5,10-methylenetetrahydrofolate + NAD+ | - |
ir | |
| 5,10-methenyltetrahydrofolate + NADH + H+ | - |
Mycolicibacterium smegmatis ATCC 700084 | 5,10-methylenetetrahydrofolate + NAD+ | - |
ir | |
| additional information | identification of the reaction product of MSMEG_6596 by LC-MS/MS | Mycolicibacterium smegmatis | ? | - |
- |
|
| additional information | identification of the reaction product of MSMEG_6649 by LC-MS/MS | Mycolicibacterium smegmatis | ? | - |
- |
|
| additional information | identification of the reaction product of MSMEG_6596 by LC-MS/MS | Mycolicibacterium smegmatis ATCC 700084 | ? | - |
- |
|
| additional information | identification of the reaction product of MSMEG_6649 by LC-MS/MS | Mycolicibacterium smegmatis ATCC 700084 | ? | - |
- |
|
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| monomer | MSMEG_6596 is a monomeric protein and lacks flavin coenzyme | Mycolicibacterium smegmatis |
| monomer | MSMEG_6649 is a monomeric protein and lacks flavin coenzyme | Mycolicibacterium smegmatis |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| MSMEG_6596 | - |
Mycolicibacterium smegmatis |
| MSMEG_6649 | - |
Mycolicibacterium smegmatis |
| MTHFR | - |
Mycolicibacterium smegmatis |
| MTHFR1 | - |
Mycolicibacterium smegmatis |
| MTHFR2 | - |
Mycolicibacterium smegmatis |
| NADH-oxidizing methylenetetrahydrofolate reductase | - |
Mycolicibacterium smegmatis |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 22 | - |
assay at room temperature | Mycolicibacterium smegmatis |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.688 | - |
5,10-methenyltetrahydrofolate | pH 7.6, 22°C | Mycolicibacterium smegmatis | |
| 1.322 | - |
NADH | pH 7.6, 22°C | Mycolicibacterium smegmatis | |
| 4.063 | - |
5,10-methenyltetrahydrofolate | pH 7.6, 22°C | Mycolicibacterium smegmatis | |
| 6.095 | - |
NADH | pH 7.6, 22°C | Mycolicibacterium smegmatis | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.6 | - |
assay at | Mycolicibacterium smegmatis |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| additional information | MSMEG_6596 is a monomeric protein and lacks flavin coenzyme. No activity with NADP(H) | Mycolicibacterium smegmatis | |
| additional information | MSMEG_6649 is a monomeric protein and lacks flavin coenzyme. No activity with NADP(H) | Mycolicibacterium smegmatis | |
| NAD+ | - |
Mycolicibacterium smegmatis | |
| NADH | - |
Mycolicibacterium smegmatis | |
General Information
| General Information | Comment | Organism |
|---|---|---|
| malfunction | the Mycobacterium smegmatis DELTAmsmeg_6596 strain is partially auxotrophic for methionine and grows only poorly without methionine or without being complemented with a functional copy of MTHFR1 or MTHFR2. Furthermore, the DELTAmsmeg_6596 strain is more sensitive to folate pathway inhibitors (sulfachloropyridazine, p-aminosalicylic acid, sulfamethoxazole, and trimethoprim) | Mycolicibacterium smegmatis |
| physiological function | MTHFR/MetF is an essential enzyme in a one-carbon metabolic pathway for de novo biosynthesis of methionine. In Mycobacterium smegmatis, the catalytic efficiency (kcat/Km) in vitro of MSMEG_6596 (MTHFR1) for 5,10-CH2-THF and NADH is 13.5 and 15.3fold higher than that of MSMEG_6649 (MTHFR2). Thus, MSMEG_6596 is the major MTHFR. This interpretation is further supported by better rescue of the Escherichia coli DELTAmthfr mutant strain by MTHFR1 than by MTHFR2. The isozymes MTHFR1 and MTHFR2 are two noncanonical MTHFR proteins that are monomeric and lack flavin coenzyme. Both MTHFR1 and MTHFR2 are involved in de novo methionine biosynthesis and required for antifolate resistance in mycobacteria. MTHFR activity is required during stress from folate pathway inhibitors. The expression of MSMEG_6596 restores maximum resistance or growth, followed by MSMEG_6649, providing antifolate resistance to Mycobacterium smegmatis | Mycolicibacterium smegmatis |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 4.75 | - |
5,10-methenyltetrahydrofolate | pH 7.6, 22°C | Mycolicibacterium smegmatis | |
| 12.24 | - |
NADH | pH 7.6, 22°C | Mycolicibacterium smegmatis | |
| 64.29 | - |
5,10-methenyltetrahydrofolate | pH 7.6, 22°C | Mycolicibacterium smegmatis | |
| 94.68 | - |
NADH | pH 7.6, 22°C | Mycolicibacterium smegmatis | |
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