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Literature summary extracted from

  • Watanabe, K.; Nureki, O.; Fukai, S.; Endo, Y.; Hori, H.
    Functional categorization of the conserved basic amino acid residues in TrmH (tRNA (Gm18) methyltransferase) enzymes (2006), J. Biol. Chem., 281, 34630-34639.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
2.1.1.34 S-adenosyl-L-methionine enhances the affinity for tRNA and induces formation of the tetramer structure Thermus thermophilus

Application

EC Number Application Comment Organism
2.1.1.34 analysis Arg-41 is the catalytic center, Lys-90, Arg-166, Arg-168, and Arg-176 are the initial sites of tRNA binding, Arg-8, Arg-19, and Lys-32 are required in the tRNA binding site for continuation the catalytic cycle, Arg-11-His-71-Met-147 interaction is involved in the structural element in release of S-adenosyl-L-homocysteine, His-34 plays a role in the assisted phosphate binding site, Arg-109 has an unknown function Thermus thermophilus

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.1.1.34 vector pET30aGm expressed in Escherichia coli Thermus thermophilus

Protein Variants

EC Number Protein Variants Comment Organism
2.1.1.34 H34A no change of affinity for S-adenosyl-L-methionine and tRNA Thermus thermophilus
2.1.1.34 H71A has increased affinity for S-adenosyl-L-methionine, decreased affinity for tRNA Thermus thermophilus
2.1.1.34 K32A no change of affinity for S-adenosyl-L-methionine, decreased affinity for tRNA Thermus thermophilus
2.1.1.34 K90A no change of affinity for S-adenosyl-L-methionine, decreased affinity for tRNA Thermus thermophilus
2.1.1.34 L179Stop no change of affinity for S-adenosyl-L-methionine, markedly decreased affinity for tRNA Thermus thermophilus
2.1.1.34 R109A no change of affinity for S-adenosyl-L-methionine and slightly increased affinity for tRNA Thermus thermophilus
2.1.1.34 R11A has lost its affinity for S-adenosyl-L-methionine, decreased affinity for tRNA Thermus thermophilus
2.1.1.34 R166A no change of affinity for S-adenosyl-L-methionine, decreased affinity for tRNA Thermus thermophilus
2.1.1.34 R168A no change of affinity for S-adenosyl-L-methionine, decreased affinity for tRNA Thermus thermophilus
2.1.1.34 R176A no change of affinity for S-adenosyl-L-methionine, decreased affinity for tRNA Thermus thermophilus
2.1.1.34 R19A no change of affinity for S-adenosyl-L-methionine, decreased affinity for tRNA Thermus thermophilus
2.1.1.34 R8A no change of affinity for S-adenosyl-L-methionine, decreased affinity for tRNA Thermus thermophilus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.1.1.34 0.017
-
 S-adenosyl-L-methionine mutant K32A Thermus thermophilus
2.1.1.34 0.019
-
 S-adenosyl-L-methionine mutant R19A Thermus thermophilus
2.1.1.34 0.02
-
 S-adenosyl-L-methionine mutant R168A Thermus thermophilus
2.1.1.34 0.021
-
 S-adenosyl-L-methionine mutant R166A Thermus thermophilus
2.1.1.34 0.021
-
 guanosine18 in tRNATyr mutant R176A Thermus thermophilus
2.1.1.34 0.022
-
 guanosine18 in tRNATyr mutant H71A Thermus thermophilus
2.1.1.34 0.023
-
 S-adenosyl-L-methionine mutant R109A Thermus thermophilus
2.1.1.34 0.024
-
 S-adenosyl-L-methionine mutant K90A Thermus thermophilus
2.1.1.34 0.024
-
 S-adenosyl-L-methionine mutant R176A Thermus thermophilus
2.1.1.34 0.044
-
 S-adenosyl-L-methionine mutant H71A Thermus thermophilus
2.1.1.34 0.045
-
 S-adenosyl-L-methionine mutant L179Stop Thermus thermophilus
2.1.1.34 0.048
-
 guanosine18 in tRNATyr mutant K90A Thermus thermophilus
2.1.1.34 0.053
-
 guanosine18 in tRNATyr mutant R166A Thermus thermophilus
2.1.1.34 0.065
-
 guanosine18 in tRNATyr mutant R168A Thermus thermophilus
2.1.1.34 0.067
-
 S-adenosyl-L-methionine mutant H34A Thermus thermophilus
2.1.1.34 0.071
-
 S-adenosyl-L-methionine mutant R8A Thermus thermophilus
2.1.1.34 0.076
-
 guanosine18 in tRNATyr mutant R109A Thermus thermophilus
2.1.1.34 0.08
-
 guanosine18 in tRNATyr mutant H34A Thermus thermophilus
2.1.1.34 0.087
-
 guanosine18 in tRNATyr mutant L179Stop Thermus thermophilus
2.1.1.34 0.1
-
 guanosine18 in tRNATyr mutant R19A Thermus thermophilus
2.1.1.34 0.12
-
 guanosine18 in tRNATyr mutant K32A Thermus thermophilus
2.1.1.34 0.27
-
 guanosine18 in tRNATyr mutant R8A Thermus thermophilus

Organism

EC Number Organism UniProt Comment Textmining
2.1.1.34 Thermus thermophilus
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.1.1.34 wild-type and mutants purified by gel filtration, mutants R11A and R166A purified by repeated ion-exchange chromatography, to homogeneity Thermus thermophilus

Storage Stability

EC Number Storage Stability Organism
2.1.1.34 -30 °C, Tris-HCl buffer, pH 7.5, 50% glycerol Thermus thermophilus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.1.1.34 S-adenosyl-L-methionine + tRNA
-
 Thermus thermophilus S-adenosyl-L-homocysteine + tRNA containing 2'-O-methylguanine
-
 ?

Subunits

EC Number Subunits Comment Organism
2.1.1.34 tetramer gel shift assay Thermus thermophilus

Synonyms

EC Number Synonyms Comment Organism
2.1.1.34 transfer RNA (Gm18) methyltransferase
-
 Thermus thermophilus
2.1.1.34 TrmH
-
 Thermus thermophilus