Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Thermotoga maritima |
Crystallization (Comment) | Organism |
---|---|
free TrmFO or bound to tetrahydrofolate-bound or glutathione, addition of GSH greatly improves the quality of the crystals, X-ray diffraction structure determinationand analysis at 2.1 A, 1.6 A, and 1.05 A resolutions, respectively | Thermotoga maritima |
Protein Variants | Comment | Organism |
---|---|---|
C223A | site-directed mutagenesis, almost inactive mutant | Thermotoga maritima |
C51A | site-directed mutagenesis, almost inactive mutant | Thermotoga maritima |
E341A | site-directed mutagenesis, the active mutant shows a decrease in both FAD binding and methylation activity compared to the wild-type enzyme | Thermotoga maritima |
H308A | site-directed mutagenesis, the mutant shows 57% of wild-type enzyme activity compared to the wild-type enzyme | Thermotoga maritima |
K282A | site-directed mutagenesis, almost inactive mutant | Thermotoga maritima |
K287A | site-directed mutagenesis, almost inactive mutant | Thermotoga maritima |
K409A | site-directed mutagenesis, the mutant shows decreased activity compared to the wild-type enzyme | Thermotoga maritima |
K410A | site-directed mutagenesis, the mutant shows decreased activity compared to the wild-type enzyme | Thermotoga maritima |
N310A | site-directed mutagenesis, the mutant shows 23% of wild-type enzyme activity compared to the wild-type enzyme | Thermotoga maritima |
R97A | site-directed mutagenesis, the mutant shows decreased activity compared to the wild-type enzyme | Thermotoga maritima |
W283A | site-directed mutagenesis, the mutant shows slightly decreased activity | Thermotoga maritima |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
5,10-methylenetetrahydrofolate + tRNA containing uridine at position 54 + FADH2 | Thermotoga maritima | - |
tetrahydrofolate + tRNA containing ribothymidine at position 54 + FAD | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermotoga maritima | Q9WZJ3 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant enzyme from Escherichia coli to homogeneity | Thermotoga maritima |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
5,10-methylenetetrahydrofolate + uracil54 in tRNA + NAD(P)H + H+ = tetrahydrofolate + 5-methyluracil54 in tRNA + NAD(P)+ | catalytic mechanism and active site structure, the methylene of MTHF is directly transferred onto U54, and then the exocyclic methylene of U54 is reduced by FADH2, overview | Thermotoga maritima |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
5,10-methylenetetrahydrofolate + tRNA containing uridine at position 54 + FADH2 | - |
Thermotoga maritima | tetrahydrofolate + tRNA containing ribothymidine at position 54 + FAD | - |
? | |
5,10-methylenetetrahydrofolate + tRNA containing uridine at position 54 + FADH2 | tetrahydrofolate is sandwiched between the flavin ring of FAD and the imidazole ring of a His residue, the transferring methylene group of MTHF is located close to the redox-active N5 atom of FAD, TrmFO-tRNA docking model and active site structure, overview | Thermotoga maritima | tetrahydrofolate + tRNA containing ribothymidine at position 54 + FAD | - |
? | |
additional information | assay method development and evaluation | Thermotoga maritima | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
folate/FAD-dependent tRNA T54 methyltransferase | - |
Thermotoga maritima |
TRMFO | - |
Thermotoga maritima |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
60 | - |
assay at | Thermotoga maritima |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | dependent on, recognition and binding structure, overview | Thermotoga maritima |