Literature summary for 2.1.1.256 extracted from

Fislage, M.; Roovers, M.; Tuszynska, I.; Bujnicki, J.M.; Droogmans, L.; Versees, W.
Crystal structures of the tRNA:m2G6 methyltransferase Trm14/TrmN from two domains of life (2012), Nucleic Acids Res., 40, 5149-5161.

Search for more literature for 2.1.1.256

Cloned(Commentary)

Cloned (Comment)	Organism
expression of the N-terminally His-tagged emzyme in in Escherichia coli strain Rosetta (DE3)	Pyrococcus furiosus
expression of the N-terminally His-tagged enzyme in in Escherichia coli strain BL21 (DE3)	Thermus thermophilus

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant detagged enzyme in complex with S-adenosyl-L-methionine or reaction product S-adenosyl-homocysteine, and inhibitor sinefungin, hanging drop vapor diffusion, protein in 50 mM Tris-HCl, pH 8.0, 10 mM MgCl2, 500 mM NaCl, 280 mM imidazole, and 1 mM DTT, mixing in a 1:1 ratio with crystallization solution containing 100 mM Tris-acetate, pH 8.0, 32% PEG 4000, and 15% glycerol, for the phasing, selenomethionine-derivatized PfTrm14 is crystallized in a crystallization solution containing 100 mM Tris-acetate, pH 8.0, 32% PEG 4000, 15% glycerol, after streak seeding from a native PfTrm14 crystal, crystal soaking in mother liquor containing 1 mM ligand, X-ray diffraction structure determination and analysis at 1.95-2.4 A resolution	Pyrococcus furiosus
purified recombinant His-tagged enzyme in complex with S-adenosyl-L-methionine or reaction product S-adenosyl-homocysteine, and inhibitor sinefungin, hanging drop vapor diffusion, protein in 50 mM Tris-HCl, pH 8.0, 250 mM NaCl, and 350 mM imidazole, mixing in a 1:1 ratio with crystallization solution 100 mM citrate/phosphate, pH 3.5, 15% PEG 6000, 200 mM NaCl, and 100 mM sodium citrate, crystal soaking in mother liquor containing 1 mM ligand, X-ray diffraction structure determination and analysis at 2.16 A resolution, solved in the apo state by molecular replacement to a resolution of 2.05 A	Thermus thermophilus

Crystallization (Comment)

Organism

purified recombinant detagged enzyme in complex with S-adenosyl-L-methionine or reaction product S-adenosyl-homocysteine, and inhibitor sinefungin, hanging drop vapor diffusion, protein in 50 mM Tris-HCl, pH 8.0, 10 mM MgCl2, 500 mM NaCl, 280 mM imidazole, and 1 mM DTT, mixing in a 1:1 ratio with crystallization solution containing 100 mM Tris-acetate, pH 8.0, 32% PEG 4000, and 15% glycerol, for the phasing, selenomethionine-derivatized PfTrm14 is crystallized in a crystallization solution containing 100 mM Tris-acetate, pH 8.0, 32% PEG 4000, 15% glycerol, after streak seeding from a native PfTrm14 crystal, crystal soaking in mother liquor containing 1 mM ligand, X-ray diffraction structure determination and analysis at 1.95-2.4 A resolution

Pyrococcus furiosus

purified recombinant His-tagged enzyme in complex with S-adenosyl-L-methionine or reaction product S-adenosyl-homocysteine, and inhibitor sinefungin, hanging drop vapor diffusion, protein in 50 mM Tris-HCl, pH 8.0, 250 mM NaCl, and 350 mM imidazole, mixing in a 1:1 ratio with crystallization solution 100 mM citrate/phosphate, pH 3.5, 15% PEG 6000, 200 mM NaCl, and 100 mM sodium citrate, crystal soaking in mother liquor containing 1 mM ligand, X-ray diffraction structure determination and analysis at 2.16 A resolution, solved in the apo state by molecular replacement to a resolution of 2.05 A

Thermus thermophilus

Inhibitors

Inhibitors	Comment	Organism	Structure
sinefungin	-	Pyrococcus furiosus
sinefungin	-	Thermus thermophilus

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	required	Pyrococcus furiosus
Mg2+	required	Thermus thermophilus

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
S-adenosyl-L-methionine + guanine6 in tRNA	Thermus thermophilus	the enzyme specifically modifies tRNAPhe at guanosine 6 in the tRNA acceptor stem	S-adenosyl-L-homocysteine + N2-methylguanine6 in tRNA	-	?
S-adenosyl-L-methionine + guanine6 in tRNA	Pyrococcus furiosus	the enzyme specifically modifies tRNAPhe at guanosine 6 in the tRNA acceptor stem	S-adenosyl-L-homocysteine + N2-methylguanine6 in tRNA	-	?

Organism

Organism	UniProt	Comment	Textmining
Pyrococcus furiosus	Q8U248	gene PF1002	-
Thermus thermophilus	Q72IH5	gene TTC1157	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
additional information	docking model of TrmN in complex with tRNAPhe of Thermus thermophilus	Thermus thermophilus	?	-	?
additional information	active site structure with bound ligands, S-adenosyl-L-methionine, S-adenosyl-L-homocysteine, and sinefungin, overview	Pyrococcus furiosus	?	-	?
S-adenosyl-L-methionine + guanine6 in tRNA	the enzyme specifically modifies tRNAPhe at guanosine 6 in the tRNA acceptor stem	Thermus thermophilus	S-adenosyl-L-homocysteine + N2-methylguanine6 in tRNA	-	?
S-adenosyl-L-methionine + guanine6 in tRNA	the enzyme specifically modifies tRNAPhe at guanosine 6 in the tRNA acceptor stem	Pyrococcus furiosus	S-adenosyl-L-homocysteine + N2-methylguanine6 in tRNA	-	?
S-adenosyl-L-methionine + guanine6 in tRNA	the enzyme specifically modifies tRNAPhe at guanosine 6 in the tRNA acceptor stem. Contribution of the THUMP domain in tRNA recognition and catalysis, substrate binding and ligand-induced conformational changes in the RFM domain, overview	Thermus thermophilus	S-adenosyl-L-homocysteine + N2-methylguanine6 in tRNA	-	?
S-adenosyl-L-methionine + guanine6 in tRNA	the enzyme specifically modifies tRNAPhe at guanosine 6 in the tRNA acceptor stem. Contribution of the THUMP domain in tRNA recognition and catalysis, substrate binding and ligand-induced conformational changes in the RFM domain	Pyrococcus furiosus	S-adenosyl-L-homocysteine + N2-methylguanine6 in tRNA	-	?

Subunits

Subunits	Comment	Organism
additional information	the enzyme consists of an N-terminal THUMP domain fused to a catalytic Rossmann-fold MTase domain, with the main tRNA binding surface in a groove between the THUMP domain and the MTase domain, secondary sequence comparison, overview	Thermus thermophilus
additional information	the enzyme consists of an N-terminal THUMP domain fused to a catalytic Rossmann-fold MTase domain, with the main tRNA binding surface in a groove between the THUMP domain and the MTase domain, secondary sequence comparison, overview	Pyrococcus furiosus

Synonyms

Synonyms	Comment	Organism
G6 MTase	-	Thermus thermophilus
G6 MTase	-	Pyrococcus furiosus
PfTrm14	-	Pyrococcus furiosus
Trm14	-	Thermus thermophilus
Trm14	-	Pyrococcus furiosus
tRNA m2G6 MTase	-	Thermus thermophilus
tRNA m2G6 MTase	-	Pyrococcus furiosus
tRNA:m2G6 methyltransferase	-	Thermus thermophilus
tRNA:m2G6 methyltransferase	-	Pyrococcus furiosus
TTCTrmN	-	Thermus thermophilus

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
60	-	assay at	Thermus thermophilus
70	-	assay at	Pyrococcus furiosus

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8	-	assay at	Thermus thermophilus
8	-	assay at	Pyrococcus furiosus

General Information

General Information	Comment	Organism
evolution	RNA MTases from the TrmN/Trm14 family are present in archaea, bacteria and eukaryota and all specifically modify tRNAPhe at guanosine 6 in the tRNA acceptor stem. RNA MTases can be classified into four superfamilies, overview	Thermus thermophilus
evolution	RNA MTases from the TrmN/Trm14 family are present in archaea, bacteria and eukaryota and all specifically modify tRNAPhe at guanosine 6 in the tRNA acceptor stem. RNA MTases can be classified into four superfamilies, overview	Pyrococcus furiosus