Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli | Arthrobacter globiformis |
Crystallization (Comment) | Organism |
---|---|
purified recombinant wild-type enzyme, sitting-drop vapour diffusion method, 0.004 ml of protein solution containing 15 mg/ml protein, mixed with 0.002 ml reservoir solution containing 15% PEG 5000 monomethylether, 0.2 M MgCl2, and 0.1 M HEPES, pH 7.5, magnesium salt can be exchanged for acetate, for complexing the crystals are soaked in a solution containing folic or folinic acid in 15% PEG 5000 monomethylether and 0.3 M NaCl for 10 min, X-ray diffraction structure determination and analysis at 1.60 A resolution | Arthrobacter globiformis |
Protein Variants | Comment | Organism |
---|---|---|
H225Q | site-directed mutagenesis, reduced catalytic activity compared to the wild-type enzyme | Arthrobacter globiformis |
Y259F | site-directed mutagenesis, reduced catalytic activity compared to the wild-type enzyme | Arthrobacter globiformis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.4 | - |
N,N-dimethylglycine | wild-type enzyme | Arthrobacter globiformis | |
47 | - |
N,N-dimethylglycine | mutant Y259F | Arthrobacter globiformis | |
59 | - |
N,N-dimethylglycine | mutant H225Q | Arthrobacter globiformis |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
mitochondrion | - |
Arthrobacter globiformis | 5739 | - |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Arthrobacter globiformis | bifunctional enzyme catalyzing the formation of 5,10-methylene tetrahydrofolate and the oxidation of N,N-dimethylglycine, folate binding and active site, a large channel connects both active sites, the channeling is kinetically controlled by terahydrofolate binding, overview | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Arthrobacter globiformis | - |
bifunctional enzyme | - |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
N,N-dimethylglycine + 5,6,7,8-tetrahydrofolate + O2 = sarcosine + 5,10-methylenetetrahydrofolate + H2O2 | reaction mechanism, substrate binding structures, bifunctional enzyme catalyzing the formation of 5,10-methylne tetrahydrofolate and the oxidation of N,N-dimethylglycine, active site structures, Tyr259 and His225 are involved in catalysis | Arthrobacter globiformis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | bifunctional enzyme catalyzing the formation of 5,10-methylene tetrahydrofolate and the oxidation of N,N-dimethylglycine, folate binding and active site, a large channel connects both active sites, the channeling is kinetically controlled by terahydrofolate binding, overview | Arthrobacter globiformis | ? | - |
? | |
N,N-dimethylglycine + 5,6,7,8-tetrahydrofolate + O2 | - |
Arthrobacter globiformis | sarcosine + 5,10-methylenetetrahydrofolate + H2O2 | - |
? | |
N,N-dimethylglycine + H2O + O2 | In the absence of tetrahydrofolate, formaldehyde is the second product of catalysis, formed by hydrolysis of a labile iminium intermediate. In the cell, toxic formaldehyde is not produced and these enzymes catalyse the formation of 5,10-methylene-THF from THF and the labile iminium intermediate. | Arthrobacter globiformis | sarcosine + formaldehyde + H2O2 | - |
? |
Synonyms | Comment | Organism |
---|---|---|
DMGO | - |
Arthrobacter globiformis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.047 | - |
N,N-dimethylglycine | mutant Y259F | Arthrobacter globiformis | |
4.35 | - |
N,N-dimethylglycine | mutant H225Q | Arthrobacter globiformis | |
10.6 | - |
N,N-dimethylglycine | wild-type enzyme | Arthrobacter globiformis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | covalently bound to the N-terminus, biding site structure | Arthrobacter globiformis |