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Literature summary for 2.1.1.35 extracted from
- Purification of transfer RNA (m5U54)-methyltransferase from Escherichia coli. Association with RNA (1988), Eur. J. Biochem., 177, 467-475.
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| dithiothreitol | activates | Escherichia coli |  |
| glutathione | activates | Escherichia coli | |
| putrescine | activates, inhibitory at optimal Mg2+ concentration | Escherichia coli | |
| spermidine | activates, inhibitory at optimal Mg2+ concentration | Escherichia coli | |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| hybrid plasmid carrying trmA gene encoding enzyme is used to amplify enzyme production, trmA gene is sequenced | Escherichia coli |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| K+ | inhibitory in presence of Mg2+ | Escherichia coli | |
| Na+ | inhibitory in presence of Mg2+ | Escherichia coli | |
| NH4+ | inhibitory in presence of Mg2+ | Escherichia coli | |
| p-chloromercuribenzoate | at 2 mM, complete inhibition | Escherichia coli | |
| putrescine | inhibition at optimal Mg2+-concentration | Escherichia coli | |
| S-adenosyl-L-homocysteine | product inhibitor | Escherichia coli | |
| spermidine | inhibition at optimal Mg2+-concentration | Escherichia coli | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.00008 | - |
uridine54 in tRNA | - |
Escherichia coli | |
| 0.017 | - |
S-adenosyl-L-methionine | - |
Escherichia coli | |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| soluble | - |
Escherichia coli | - |
- |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Mg2+ | activates at 10 mM | Escherichia coli | |
| additional information | no require of cations | Escherichia coli |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 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 |
| 54000 | - |
RNA-enzyme complex: RNA component associated with the 41/42 kDa polypeptide, PAGE | Escherichia coli |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| S-adenosyl-L-methionine + uridine54 in tRNA | Escherichia coli | tRNA modifying enzyme | S-adenosyl-L-homocysteine + 5-methyluridine54 in tRNA | - |
? | |
| 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 | - |
? | |
| S-adenosyl-L-methionine + uridine54 in tRNA | Escherichia coli C600 / pTN106 | tRNA modifying enzyme | S-adenosyl-L-homocysteine + 5-methyluridine54 in tRNA | - |
? | |
| 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
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Escherichia coli | - |
- |
- |
| Escherichia coli | - |
native enzyme, may be in 2 forms of 41 and 42 kDa, and TrmA/RNA complex | - |
| Escherichia coli C600 / pTN106 | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| 5800fold purification | Escherichia coli |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 0.46 | - |
- |
Escherichia coli |
Storage Stability
| Storage Stability | Organism |
|---|---|
| 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)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 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 | - |
? | |
| 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 | - |
? | |
| 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 | ? | |
| 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 | ? | |
| S-adenosyl-L-methionine + uridine54 in tRNA | tRNA modifying enzyme | Escherichia coli | S-adenosyl-L-homocysteine + 5-methyluridine54 in tRNA | - |
? | |
| 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 | - |
? | |
| 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 | ? | |
| 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 | ? | |
| S-adenosyl-L-methionine + uridine54 in tRNA | tRNA modifying enzyme | Escherichia coli C600 / pTN106 | S-adenosyl-L-homocysteine + 5-methyluridine54 in tRNA | - |
? | |
| 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
| Subunits | Comment | Organism |
|---|---|---|
| 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]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | - |
assay at | Escherichia coli |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | - |
2 h, stable | Escherichia coli |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.317 | - |
uridine54 in tRNA | - |
Escherichia coli | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| additional information | - |
pI: 4.7 | Escherichia coli |
| 8 | - |
- |
Escherichia coli |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| S-adenosyl-L-methionine | uses S-adenosyl-L-methionine as cofactor | Escherichia coli | |
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