Literature summary for 2.1.1.74 extracted from

Nishimasu, H.; Ishitani, R.; Yamashita, K.; Iwashita, C.; Hirata, A.; Hori, H.; Nureki, O.
Atomic structure of a folate/FAD-dependent tRNA T54 methyltransferase (2009), Proc. Natl. Acad. Sci. USA, 106, 8180-8185.

Search for more literature for 2.1.1.74

Cloned(Commentary)

Cloned (Comment)	Organism
expression in Escherichia coli	Thermotoga maritima

Crystallization (Commentary)

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

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/ Products (Substrates)

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

Organism	UniProt	Comment	Textmining
Thermotoga maritima	Q9WZJ3	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant enzyme from Escherichia coli to homogeneity	Thermotoga maritima

Reaction

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	a

Substrates and Products (Substrate)

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

Synonyms	Comment	Organism
folate/FAD-dependent tRNA T54 methyltransferase	-	Thermotoga maritima
TRMFO	-	Thermotoga maritima

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
60	-	assay at	Thermotoga maritima

Cofactor

Cofactor	Comment	Organism	Structure
FAD	dependent on, recognition and binding structure, overview	Thermotoga maritima

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Release 2023.1 (January 2023)