Cloned (Comment) | Organism |
---|---|
overexpression of His-tagged enzyme in Escherichia coli strain BL21(DE3) | Mycoplasma pneumoniae |
Crystallization (Comment) | Organism |
---|---|
purified recombinant enzyme complexed with ADP, phosphate, and 5-formyltetrahydrofolate, hanging drop vapour diffusion method, 20°C, 30 mg/ml protein with 20% PEG 4000, 5 mM ATP, and 10 mM 5-formylTHF, flash freezing of all crystals in 20% PEG 4000 and 30% ethylene glycol, X-ray diffraction structure determination and analysis at 2.5 A resolution, crystallization of the enzyme with ATP analogues ATP-gamma-S, AMP-PNP, or AMP-PCP is not successful | Mycoplasma pneumoniae |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
phosphate | product inhibition | Mycoplasma pneumoniae |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | steady-state kinetics, recombinant enzyme | Mycoplasma pneumoniae | |
0.165 | - |
5-formyltetrahydrofolate | pH 6.0, 37°C, recombinant enzyme | Mycoplasma pneumoniae | |
0.166 | - |
ATP | pH 6.0, 37°C, recombinant enzyme | Mycoplasma pneumoniae |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | activates, can substitute for Mg2+ | Mycoplasma pneumoniae | |
Co2+ | activates, can substitute for Mg2+ | Mycoplasma pneumoniae | |
Cu2+ | activates, can substitute for Mg2+ | Mycoplasma pneumoniae | |
Fe2+ | activates, can substitute for Mg2+ | Mycoplasma pneumoniae | |
Mg2+ | dependent on, most effective divalent cation, required for MgATP2- complex formation, binding structure | Mycoplasma pneumoniae | |
Mn2+ | activates, can substitute for Mg2+ | Mycoplasma pneumoniae | |
additional information | the enzyme is dependent on divalent cation, Mg2+ is preferred | Mycoplasma pneumoniae | |
Zn2+ | activates, can substitute for Mg2+ | Mycoplasma pneumoniae |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
22000 | - |
monomeric recombinant enzyme, gel filtration | Mycoplasma pneumoniae |
44000 | - |
dimeric recombinant enzyme, gel filtration | Mycoplasma pneumoniae |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + 5-formyltetrahydrofolate | Mycoplasma pneumoniae | - |
ADP + 5,10-methenyltetrahydrofolate + phosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mycoplasma pneumoniae | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by metal affinity chromatography to over 95% homogeneity | Mycoplasma pneumoniae |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
ATP + 5-formyltetrahydrofolate = ADP + phosphate + 5,10-methenyltetrahydrofolate | active site and substrate binding site structure, binding and catalytic mechanism | Mycoplasma pneumoniae |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
2.8 | - |
purified recombinant enzyme | Mycoplasma pneumoniae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + 5-formyltetrahydrofolate | ATP in form of MgATP2- | Mycoplasma pneumoniae | ADP + phosphate + 5,10-methenyltetrahydrofolate | - |
? | |
ATP + 5-formyltetrahydrofolate | - |
Mycoplasma pneumoniae | ADP + 5,10-methenyltetrahydrofolate + phosphate | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | 2 * 22000, recombinant enzyme, SDS-PAGE, the recombinant enyme exists as a mixture of monomers and dimers | Mycoplasma pneumoniae |
monomer | 1 * 22000, recombinant enzyme, SDS-PAGE, the recombinant enyme exists as a mixture of monomers and dimers | Mycoplasma pneumoniae |
Synonyms | Comment | Organism |
---|---|---|
5,10-Methenyltetrahydrofolate synthetase | - |
Mycoplasma pneumoniae |
Methenyltetrahydrofolate synthetase | - |
Mycoplasma pneumoniae |
More | the enzyme belongs to the cycloligase protein family | Mycoplasma pneumoniae |
MTHFS | - |
Mycoplasma pneumoniae |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Mycoplasma pneumoniae |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.94 | - |
5-formyltetrahydrofolate | pH 6.0, 37°C, recombinant enzyme | Mycoplasma pneumoniae | |
0.99 | - |
ATP | pH 6.0, 37°C, recombinant enzyme | Mycoplasma pneumoniae |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | - |
assay at | Mycoplasma pneumoniae |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | absolutely required, as MgATP2-, binding structure | Mycoplasma pneumoniae | |
additional information | no activity with nucleoside 5'-mono- or 5'-diphosphates | Mycoplasma pneumoniae |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
2.12 | - |
phosphate | pH 6.0, 37°C, recombinant enzyme | Mycoplasma pneumoniae |