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 (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 | Comment | Organism | Structure |
---|---|---|---|
sinefungin | - |
Pyrococcus furiosus | |
sinefungin | - |
Thermus thermophilus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Thermus thermophilus | |
Mg2+ | required | Pyrococcus furiosus |
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 | UniProt | Comment | Textmining |
---|---|---|---|
Pyrococcus furiosus | Q8U248 | gene PF1002 | - |
Thermus thermophilus | Q72IH5 | gene TTC1157 | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | active site structure with bound ligands, S-adenosyl-L-methionine, S-adenosyl-L-homocysteine, and sinefungin, overview | Pyrococcus furiosus | ? | - |
? | |
additional information | docking model of TrmN in complex with tRNAPhe of Thermus thermophilus | Thermus thermophilus | ? | - |
? | |
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 | 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 | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | 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 |
More | 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 | 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 Maximum [°C] | Comment | Organism |
---|---|---|---|
60 | - |
assay at | Thermus thermophilus |
70 | - |
assay at | Pyrococcus furiosus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Thermus thermophilus |
8 | - |
assay at | Pyrococcus furiosus |
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 |