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Literature summary for 2.1.1.228 extracted from

  • O'Dwyer, K.; Watts, J.M.; Biswas, S.; Ambrad, J.; Barber, M.; Brule, H.; Petit, C.; Holmes, D.J.; Zalacain, M.; Holmes, W.M.
    Characterization of Streptococcus pneumoniae TrmD, a tRNA methyltransferase essential for growth (2004), J. Bacteriol., 186, 2346-2354.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli by using a His-tagged T7 expression vector Streptococcus pneumoniae

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0007
-
S-adenosyl-L-homocysteine pH 8.5, 37°C Streptococcus pneumoniae
0.0043
-
guanine37 in tRNALeu(GAC) pH 8.5, 37°C Streptococcus pneumoniae

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ divalent cation (in oder of decreasing efficiency: Mg2+, Mn2+, Ca2+) required, optimal activity in presence of 10 mM Mg2+ Streptococcus pneumoniae
Mg2+ divalent cation (in oder of decreasig efficiency: Mg2+, Mn2+, Ca2+) required, optimal activity in presence of 10 mM Mg2+ Streptococcus pneumoniae
Mn2+ divalent cation (in oder of decreasing efficiency: Mg2+, Mn2+, Ca2+) required, optimal activity in presence of 10 mM Mg2+ Streptococcus pneumoniae
additional information no activity is observed in absence of divalent cation or ion presence of Co2+. TrmD activity increases only 10% in the presence of a monovalent cation, indicating the nonessential nature of this factor Streptococcus pneumoniae

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
32000
-
2 * 32000 Streptococcus pneumoniae
65000
-
dynamic light scattering Streptococcus pneumoniae
69000
-
gel filtration Streptococcus pneumoniae

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
S-adenosyl-L-methionine + guanine37 in tRNA Streptococcus pneumoniae
-
S-adenosyl-L-homocysteine + N1-methylguanine37 in tRNA
-
?

Organism

Organism UniProt Comment Textmining
Streptococcus pneumoniae
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant protein Streptococcus pneumoniae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
S-adenosyl-L-methionine + guanine37 in tRNA
-
Streptococcus pneumoniae S-adenosyl-L-homocysteine + N1-methylguanine37 in tRNA
-
?
S-adenosyl-L-methionine + guanine37 in tRNA the streptococcal enzyme utilizes a sequential mechanism. Nonsubstrate tRNA species, like tRNAThr(GGT), tRNAPhe, and tRNAAla(TGC), bind the enzyme with similar affinities, suggesting that tRNA specificity is achieved via a postbinding events. The streptococcal TrmD requires the complete tRNA structure since it cannot modify the tRNALeu(CAG) minihelix lacking the D, T, and extra loops of complete tRNA. In addition, and consistent with a requirement for G at positions 36 and 37 in the tRNA, the enzyme methylates yeast tRNAAsp possessing a G36G37 sequence with kinetic values that are indistinguishable from those obtained with substrate tRNALeu(CAG) but does not methylate either tRNAAsp possessing a C36G37 sequence or tRNA(Asp) possessing a C36A37 sequence Streptococcus pneumoniae S-adenosyl-L-homocysteine + N1-methylguanine37 in tRNA
-
?
S-adenosyl-L-methionine + guanine37 in tRNALeu(GAC)
-
Streptococcus pneumoniae S-adenosyl-L-homocysteine + N1-methylguanine37 in tRNALeu(GAC)
-
?

Subunits

Subunits Comment Organism
dimer 2 * 32000 Streptococcus pneumoniae

Synonyms

Synonyms Comment Organism
TrmD
-
Streptococcus pneumoniae

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Streptococcus pneumoniae

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.25
-
S-adenosyl-L-homocysteine pH 8.5, 37°C Streptococcus pneumoniae
0.25
-
guanine37 in tRNALeu(GAC) pH 8.5, 37°C Streptococcus pneumoniae

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7 8
-
Streptococcus pneumoniae
8.5
-
assay at Streptococcus pneumoniae

pH Range

pH Minimum pH Maximum Comment Organism
6 8.5 pH 6.0: about 50% of maximal activity, pH 8.5: about 75% of maximal activity Streptococcus pneumoniae

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
58
-
S-adenosyl-L-homocysteine pH 8.5, 37°C Streptococcus pneumoniae
357
-
guanine37 in tRNALeu(GAC) pH 8.5, 37°C Streptococcus pneumoniae