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Literature summary extracted from

  • Ny, T.; Lindström, P.H.R.; Hagervall, T.G.; Björk, G.R.
    Purification of transfer RNA (m5U54)-methyltransferase from Escherichia coli. Association with RNA (1988), Eur. J. Biochem., 177, 467-475.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
2.1.1.35 dithiothreitol activates Escherichia coli
2.1.1.35 glutathione activates Escherichia coli
2.1.1.35 putrescine activates, inhibitory at optimal Mg2+ concentration Escherichia coli
2.1.1.35 spermidine activates, inhibitory at optimal Mg2+ concentration Escherichia coli

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.1.1.35 hybrid plasmid carrying trmA gene encoding enzyme is used to amplify enzyme production, trmA gene is sequenced Escherichia coli

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.1.1.35 K+ inhibitory in presence of Mg2+ Escherichia coli
2.1.1.35 Na+ inhibitory in presence of Mg2+ Escherichia coli
2.1.1.35 NH4+ inhibitory in presence of Mg2+ Escherichia coli
2.1.1.35 p-chloromercuribenzoate at 2 mM, complete inhibition Escherichia coli
2.1.1.35 putrescine inhibition at optimal Mg2+-concentration Escherichia coli
2.1.1.35 S-adenosyl-L-homocysteine product inhibitor Escherichia coli
2.1.1.35 spermidine inhibition at optimal Mg2+-concentration Escherichia coli

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.1.1.35 0.00008
-
 uridine54 in tRNA
-
 Escherichia coli
2.1.1.35 0.017
-
 S-adenosyl-L-methionine
-
 Escherichia coli

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
2.1.1.35 soluble
-
 Escherichia coli
-
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.1.1.35 Mg2+ activates at 10 mM Escherichia coli
2.1.1.35 additional information no require of cations Escherichia coli

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.1.1.35 42000
-
 1 * 42000, native protein, additional bands at 32, 41, 48-57 kDa, polypeptides ranging over 48-57 kDa are dependent on the gel system used and refer to the RNA-enzyme complex, SDS-PAGE Escherichia coli
2.1.1.35 54000
-
 RNA-enzyme complex: RNA component associated with the 41/42 kDa polypeptide, PAGE Escherichia coli

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.1.1.35 S-adenosyl-L-methionine + uridine54 in tRNA Escherichia coli tRNA modifying enzyme S-adenosyl-L-homocysteine + 5-methyluridine54 in tRNA
-
 ?
2.1.1.35 S-adenosyl-L-methionine + uridine54 in tRNA Escherichia coli involved in biosynthesis of all tRNA chains, association of enzyme to tRNA may be of regulatory significance S-adenosyl-L-homocysteine + 5-methyluridine54 in tRNA
-
 ?
2.1.1.35 S-adenosyl-L-methionine + uridine54 in tRNA Escherichia coli C600 / pTN106 tRNA modifying enzyme S-adenosyl-L-homocysteine + 5-methyluridine54 in tRNA
-
 ?
2.1.1.35 S-adenosyl-L-methionine + uridine54 in tRNA Escherichia coli C600 / pTN106 involved in biosynthesis of all tRNA chains, association of enzyme to tRNA may be of regulatory significance S-adenosyl-L-homocysteine + 5-methyluridine54 in tRNA
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.1.1.35 Escherichia coli
-
-
-
2.1.1.35 Escherichia coli
-
 native enzyme, may be in 2 forms of 41 and 42 kDa, and TrmA/RNA complex
-
2.1.1.35 Escherichia coli C600 / pTN106
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.1.1.35 5800fold purification Escherichia coli

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.1.1.35 0.46
-
-
 Escherichia coli

Storage Stability

EC Number Storage Stability Organism
2.1.1.35 4°C, crude extract, 10 mM potassium phosphate, pH 7, 1 mM EDTA, 0.1 mM dithiothreitol, 10% v/v ethylene glycol, 0.02% sodium azide, 7 days, 10% loss of activity Escherichia coli

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.1.1.35 additional information enzyme is present as 42 kDa native protein or as 48-57 kDa RNA-protein complex, RNA is not essential, but may increase enzyme activity Escherichia coli additional information
-
 ?
2.1.1.35 additional information enzyme is present as 42 kDa native protein or as 48-57 kDa RNA-protein complex, RNA is not essential, but may increase enzyme activity Escherichia coli C600 / pTN106 additional information
-
 ?
2.1.1.35 S-adenosyl-L-methionine + uridine54 in tRNA methyl group acceptor: m5U-deficient tRNA from E. coli GB1-5-39 Escherichia coli S-adenosyl-L-homocysteine + 5-methyluridine54 in tRNA formation of 5-methyluridine in position 54 of tRNA, in the TPsiC sequence in loop IV ?
2.1.1.35 S-adenosyl-L-methionine + uridine54 in tRNA enzyme is normally tightly associated with the tRNA Escherichia coli S-adenosyl-L-homocysteine + 5-methyluridine54 in tRNA formation of 5-methyluridine in position 54 of tRNA, in the TPsiC sequence in loop IV ?
2.1.1.35 S-adenosyl-L-methionine + uridine54 in tRNA tRNA modifying enzyme Escherichia coli S-adenosyl-L-homocysteine + 5-methyluridine54 in tRNA
-
 ?
2.1.1.35 S-adenosyl-L-methionine + uridine54 in tRNA involved in biosynthesis of all tRNA chains, association of enzyme to tRNA may be of regulatory significance Escherichia coli S-adenosyl-L-homocysteine + 5-methyluridine54 in tRNA
-
 ?
2.1.1.35 S-adenosyl-L-methionine + uridine54 in tRNA methyl group acceptor: m5U-deficient tRNA from E. coli GB1-5-39 Escherichia coli C600 / pTN106 S-adenosyl-L-homocysteine + 5-methyluridine54 in tRNA formation of 5-methyluridine in position 54 of tRNA, in the TPsiC sequence in loop IV ?
2.1.1.35 S-adenosyl-L-methionine + uridine54 in tRNA enzyme is normally tightly associated with the tRNA Escherichia coli C600 / pTN106 S-adenosyl-L-homocysteine + 5-methyluridine54 in tRNA formation of 5-methyluridine in position 54 of tRNA, in the TPsiC sequence in loop IV ?
2.1.1.35 S-adenosyl-L-methionine + uridine54 in tRNA tRNA modifying enzyme Escherichia coli C600 / pTN106 S-adenosyl-L-homocysteine + 5-methyluridine54 in tRNA
-
 ?
2.1.1.35 S-adenosyl-L-methionine + uridine54 in tRNA involved in biosynthesis of all tRNA chains, association of enzyme to tRNA may be of regulatory significance Escherichia coli C600 / pTN106 S-adenosyl-L-homocysteine + 5-methyluridine54 in tRNA
-
 ?

Subunits

EC Number Subunits Comment Organism
2.1.1.35 monomer 1 * 42000, native protein, additional bands at 32, 41, 48-57 kDa, polypeptides ranging over 48-57 kDa are dependent on the gel system used and refer to the RNA-enzyme complex, SDS-PAGE Escherichia coli

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.1.1.35 37
-
 assay at Escherichia coli

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.1.1.35 37
-
 2 h, stable Escherichia coli

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.1.1.35 0.317
-
 uridine54 in tRNA
-
 Escherichia coli

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.1.1.35 additional information
-
 pI: 4.7 Escherichia coli
2.1.1.35 8
-
-
 Escherichia coli

Cofactor

EC Number Cofactor Comment Organism Structure
2.1.1.35 S-adenosyl-L-methionine uses S-adenosyl-L-methionine as cofactor Escherichia coli