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Literature summary for 2.1.1.34 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
Search for more literature for 2.1.1.34

Activating Compound

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

Application

Application Comment Organism
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)

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

Protein Variants

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

KM Value [mM]

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

Organism

Organism UniProt Comment Textmining
Thermus thermophilus
-
-
-

Purification (Commentary)

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

Storage Stability

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

Substrates and Products (Substrate)

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

Subunits

Subunits Comment Organism
tetramer gel shift assay Thermus thermophilus

Synonyms

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