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Literature summary 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)

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

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.1.1.228 0.0007
-
 S-adenosyl-L-homocysteine pH 8.5, 37°C Streptococcus pneumoniae
2.1.1.228 0.0043
-
 guanine37 in tRNALeu(GAC) pH 8.5, 37°C Streptococcus pneumoniae

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.1.1.228 Ca2+ divalent cation (in oder of decreasing efficiency: Mg2+, Mn2+, Ca2+) required, optimal activity in presence of 10 mM Mg2+ Streptococcus pneumoniae
2.1.1.228 Mg2+ divalent cation (in oder of decreasig efficiency: Mg2+, Mn2+, Ca2+) required, optimal activity in presence of 10 mM Mg2+ Streptococcus pneumoniae
2.1.1.228 Mn2+ divalent cation (in oder of decreasing efficiency: Mg2+, Mn2+, Ca2+) required, optimal activity in presence of 10 mM Mg2+ Streptococcus pneumoniae
2.1.1.228 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]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.1.1.228 32000
-
 2 * 32000 Streptococcus pneumoniae
2.1.1.228 65000
-
 dynamic light scattering Streptococcus pneumoniae
2.1.1.228 69000
-
 gel filtration Streptococcus pneumoniae

Natural Substrates/ Products (Substrates)

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

Organism

EC Number Organism UniProt Comment Textmining
2.1.1.228 Streptococcus pneumoniae
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.1.1.228 recombinant protein Streptococcus pneumoniae

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.1.1.228 S-adenosyl-L-methionine + guanine37 in tRNA
-
 Streptococcus pneumoniae S-adenosyl-L-homocysteine + N1-methylguanine37 in tRNA
-
 ?
2.1.1.228 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
-
 ?
2.1.1.228 S-adenosyl-L-methionine + guanine37 in tRNALeu(GAC)
-
 Streptococcus pneumoniae S-adenosyl-L-homocysteine + N1-methylguanine37 in tRNALeu(GAC)
-
 ?

Subunits

EC Number Subunits Comment Organism
2.1.1.228 dimer 2 * 32000 Streptococcus pneumoniae

Synonyms

EC Number Synonyms Comment Organism
2.1.1.228 TrmD
-
 Streptococcus pneumoniae

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.1.1.228 37
-
 assay at Streptococcus pneumoniae

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.1.1.228 0.25
-
 S-adenosyl-L-homocysteine pH 8.5, 37°C Streptococcus pneumoniae
2.1.1.228 0.25
-
 guanine37 in tRNALeu(GAC) pH 8.5, 37°C Streptococcus pneumoniae

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.1.1.228 7 8
-
 Streptococcus pneumoniae
2.1.1.228 8.5
-
 assay at Streptococcus pneumoniae

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.1.1.228 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]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.1.1.228 58
-
 S-adenosyl-L-homocysteine pH 8.5, 37°C Streptococcus pneumoniae
2.1.1.228 357
-
 guanine37 in tRNALeu(GAC) pH 8.5, 37°C Streptococcus pneumoniae