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Literature summary for 2.1.1.221 extracted from
- Mechanistic features of the atypical tRNA m1G9 SPOUT methyltransferase, Trm10 (2017), Nucleic Acids Res., 45, 9019-9029 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene Trm10, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli | Saccharomyces cerevisiae |
| gene TrmT10A, recombinant expression of His-tagged enzyme in Escherichia coli | Homo sapiens |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| D210A | site-directed mutagenesis, the mutant does not abolish the mutant enzyme's activity, but modestly decreases it | Saccharomyces cerevisiae |
| D210K | site-directed mutagenesis, the mutant does not abolish the mutant enzyme's activity, but modestly decreases it | Saccharomyces cerevisiae |
| D210N | site-directed mutagenesis, the mutant does not abolish the mutant enzyme's activity, but modestly decreases it | Saccharomyces cerevisiae |
| D210N | site-directed mutagenesis, the mutant is resistant to 5-fluorouracil similarly to the wild-type enzyme | Homo sapiens |
| G206R | site-directed mutagenesis, inactive variant of hTRMT10A with abolished methylation activity, likely due to an inability to bind cofactor SAM | Homo sapiens |
| additional information | 5-fluorouracil (5FU) hypersensitive phenotype of trm10 deletion in Saccharomyces cerevisiae. Growth of the trm10DELTA strain is inhibited compared to a TRM10 wild-type strain in media containing 0.001 mg/ml 5FU. This growth defect is due to the loss of TRM10, since expression of a wild-type copy of either ScTrm10 or hTRMT10A complements this phenotype at either 30 °C or 37°C, while expression of the empty vector does not restore wild-type growth to the trm10DELTA strain. Cells expressing the hTRMT10A D210N variant are similarly resistant to the effects of 5FU as either wild-type enzyme, consistent with the in vitro biochemical results | Homo sapiens |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | kinetic analysis of wild-type and mutant Trm10s, overview | Saccharomyces cerevisiae | |
| additional information | - |
additional information | kinetic analysis of wild-type and mutant Trm10s, overview | Homo sapiens |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Mg2+ | activates | Saccharomyces cerevisiae |  |
| Mg2+ | activates | Homo sapiens | |
| additional information | Trm10 does not depend on a catalytic metal ion | Saccharomyces cerevisiae | |
| additional information | Trm10 does not depend on a catalytic metal ion | Homo sapiens |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| S-adenosyl-L-methionine + guanine9 in tRNA | Saccharomyces cerevisiae | - |
S-adenosyl-L-homocysteine + N1-methylguanine9 in tRNA | - |
? | |
| S-adenosyl-L-methionine + guanine9 in tRNA | Homo sapiens | - |
S-adenosyl-L-homocysteine + N1-methylguanine9 in tRNA | - |
? | |
| S-adenosyl-L-methionine + guanine9 in tRNA | Saccharomyces cerevisiae ATCC 204508 | - |
S-adenosyl-L-homocysteine + N1-methylguanine9 in tRNA | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Homo sapiens | Q8TBZ6 | - |
- |
| Saccharomyces cerevisiae | Q12400 | - |
- |
| Saccharomyces cerevisiae ATCC 204508 | Q12400 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged enzyme from Escherichia coli by nickel affinity chromatography and dialysis, to 75-90% purity | Homo sapiens |
| recombinant His-tagged wild-type and mutant enzymes from Escherichia coli by nickel affinity chromatography and dialysis, to 75-90% purity | Saccharomyces cerevisiae |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| S-adenosyl-L-methionine + guanine9 in tRNA | - |
Saccharomyces cerevisiae | S-adenosyl-L-homocysteine + N1-methylguanine9 in tRNA | - |
? | |
| S-adenosyl-L-methionine + guanine9 in tRNA | - |
Homo sapiens | S-adenosyl-L-homocysteine + N1-methylguanine9 in tRNA | - |
? | |
| S-adenosyl-L-methionine + guanine9 in tRNA | - |
Saccharomyces cerevisiae ATCC 204508 | S-adenosyl-L-homocysteine + N1-methylguanine9 in tRNA | - |
? | |
| S-adenosyl-L-methionine + guanine9 in tRNAGly | tRNA from Saccharomyces cerevisiae | Saccharomyces cerevisiae | S-adenosyl-L-homocysteine + N1-methylguanine9 in tRNAGly | - |
? | |
| S-adenosyl-L-methionine + guanine9 in tRNAGly | tRNA from Saccharomyces cerevisiae | Homo sapiens | S-adenosyl-L-homocysteine + N1-methylguanine9 in tRNAGly | - |
? | |
| S-adenosyl-L-methionine + guanine9 in tRNAGly | tRNA from Saccharomyces cerevisiae | Saccharomyces cerevisiae ATCC 204508 | S-adenosyl-L-homocysteine + N1-methylguanine9 in tRNAGly | - |
? | |
| S-adenosyl-L-methionine + guanine9 in tRNAPhe(G9) | tRNA from Saccharomyces cerevisiae | Saccharomyces cerevisiae | S-adenosyl-L-homocysteine + N1-methylguanine9 in tRNAPhe(G9) | - |
? | |
| S-adenosyl-L-methionine + guanine9 in tRNAPhe(G9) | tRNA from Saccharomyces cerevisiae | Homo sapiens | S-adenosyl-L-homocysteine + N1-methylguanine9 in tRNAPhe(G9) | - |
? | |
| S-adenosyl-L-methionine + guanine9 in tRNAPhe(G9) | tRNA from Saccharomyces cerevisiae | Saccharomyces cerevisiae ATCC 204508 | S-adenosyl-L-homocysteine + N1-methylguanine9 in tRNAPhe(G9) | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| hTRMT10A | - |
Homo sapiens |
| ScTrm10 | - |
Saccharomyces cerevisiae |
| Trm10 | - |
Saccharomyces cerevisiae |
| Trm10 | - |
Homo sapiens |
| tRNA m1G9 SPOUT methyltransferase | - |
Saccharomyces cerevisiae |
| tRNA m1G9 SPOUT methyltransferase | - |
Homo sapiens |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | - |
assay at | Saccharomyces cerevisiae |
| 30 | - |
assay at | Homo sapiens |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 8 | - |
assay at | Saccharomyces cerevisiae |
| 8 | - |
assay at | Homo sapiens |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| S-adenosyl-L-methionine | - |
Saccharomyces cerevisiae | |
| S-adenosyl-L-methionine | - |
Homo sapiens | |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | tRNA m1G9 methyltransferase (Trm10) is a member of the SpoU-TrmD (SPOUT) superfamily of methyltransferases, and Trm10 homologs are widely conserved throughout eukarya and archaea. Despite possessing the trefoil knot characteristic of SPOUT enzymes, Trm10 does not share the same quaternary structure or key sequences with other members of the SPOUT family, suggesting a distinct mechanism of catalysis. Sequence comparison of human TRMT10A and yeast Trm10. Trm10 does not depend on a catalytic metal ion, further distinguishing it from the other known SPOUT m1G methyltransferase, TrmD | Saccharomyces cerevisiae |
| evolution | tRNA m1G9 methyltransferase (Trm10) is a member of the SpoU-TrmD (SPOUT) superfamily of methyltransferases, and Trm10 homologs are widely conserved throughout eukarya and archaea. Despite possessing the trefoil knot characteristic of SPOUT enzymes, Trm10 does not share the same quaternary structure or key sequences with other members of the SPOUT family, suggesting a distinct mechanism of catalysis. Trm10 does not depend on a catalytic metal ion, further distinguishing it from the other known SPOUT m1G methyltransferase, TrmD | Homo sapiens |
| additional information | the proposed aspartate general base D210 is not critical for methylation activity, mechanism of m1G9 methylation by Trm10. The pH-rate analysis suggests that D210 and other conserved carboxylate-containing residues at the active site collaborate to establish an active site environment that promotes a single ionization that is required for catalysis. Active site residues structure-function analysis | Saccharomyces cerevisiae |
| additional information | the proposed aspartate general base D210 is not critical for methylation activity, mechanism of m1G9 methylation by Trmt10A. The pH-rate analysis suggests that D210 and other conserved carboxylate-containing residues at the active site collaborate to establish an active site environment that promotes a single ionization that is required for catalysis. Active site residues structure-function analysis | Homo sapiens |
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