Cloned (Comment) | Organism |
---|---|
recombinant expression of C-terminally His6-tagged enzyme | Pyrococcus abyssi |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | steady-state fluorescent assay and stopped-flow kinetics with 2-aminopurine-modifed tRNA substrates, , overview | Pyrococcus abyssi |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Pyrococcus abyssi |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
2 S-adenosyl-L-methionine + adenine57/adenine58 in tRNA | Pyrococcus abyssi | the solvent accessibility of the S-adenosyl-L-methionine pocket is not affected by the tRNA, thereby enabling S-adenosyl-L-homocysteine to be replaced by S-adenosyl-L-methionine without prior release of monomethylated tRNA | 2 S-adenosyl-L-homocysteine + N1-methyladenine57/N1-methyladenine58 in tRNA | - |
? | |
2 S-adenosyl-L-methionine + adenine57/adenine58 in tRNA | Pyrococcus abyssi GE5 / CNCM I-1302 / DSM 25543 | the solvent accessibility of the S-adenosyl-L-methionine pocket is not affected by the tRNA, thereby enabling S-adenosyl-L-homocysteine to be replaced by S-adenosyl-L-methionine without prior release of monomethylated tRNA | 2 S-adenosyl-L-homocysteine + N1-methyladenine57/N1-methyladenine58 in tRNA | - |
? | |
additional information | Pyrococcus abyssi | in most organisms, the widely conserved 1-methyladenosine58 (m1A58) tRNA modification is catalyzed by S-adenosyl-L-methionine-dependent site-specific enzyme TrmI. In archaea, TrmI also methylates the adjacent adenine 57, m1A57 being an obligatory intermediate of 1-methyl-inosine57 formation, multi-site specificity mechanism, overview | ? | - |
? | |
additional information | Pyrococcus abyssi GE5 / CNCM I-1302 / DSM 25543 | in most organisms, the widely conserved 1-methyladenosine58 (m1A58) tRNA modification is catalyzed by S-adenosyl-L-methionine-dependent site-specific enzyme TrmI. In archaea, TrmI also methylates the adjacent adenine 57, m1A57 being an obligatory intermediate of 1-methyl-inosine57 formation, multi-site specificity mechanism, overview | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pyrococcus abyssi | Q9V1J7 | - |
- |
Pyrococcus abyssi | Q9V1J7 | gene PAB0283 or trmI | - |
Pyrococcus abyssi GE5 / CNCM I-1302 / DSM 25543 | Q9V1J7 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Pyrococcus abyssi |
recombinant C-terminally His6-tagged enzyme by nickel affiniyt chromatography and gel filtration | Pyrococcus abyssi |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2 S-adenosyl-L-methionine + adenine57/adenine58 in tRNA | the solvent accessibility of the S-adenosyl-L-methionine pocket is not affected by the tRNA, thereby enabling S-adenosyl-L-homocysteine to be replaced by S-adenosyl-L-methionine without prior release of monomethylated tRNA | Pyrococcus abyssi | 2 S-adenosyl-L-homocysteine + N1-methyladenine57/N1-methyladenine58 in tRNA | - |
? | |
2 S-adenosyl-L-methionine + adenine57/adenine58 in tRNA | the solvent accessibility of the S-adenosyl-L-methionine pocket is not affected by the tRNA, thereby enabling S-adenosyl-L-homocysteine to be replaced by S-adenosyl-L-methionine without prior release of monomethylated tRNA | Pyrococcus abyssi GE5 / CNCM I-1302 / DSM 25543 | 2 S-adenosyl-L-homocysteine + N1-methyladenine57/N1-methyladenine58 in tRNA | - |
? | |
2 S-adenosyl-L-methionine + adenine57/adenine58 in tRNAArg | modified mini-tRNAAsp substrate, PabTrmI recognizes and methylates only A58 in mini-tRNAArgTCT containing the G57A58A59U60 sequence | Pyrococcus abyssi | 2 S-adenosyl-L-homocysteine + adenine57/N1-methyladenine58 in tRNAArg | - |
? | |
2 S-adenosyl-L-methionine + adenine57/adenine58 in tRNAAsp | modified mini-tRNAAsp substrate, PabTrmI recognizes and methylates both A57 and A58 in mini-tRNAAsp containing the A57A58A59U60 sequence | Pyrococcus abyssi | 2 S-adenosyl-L-homocysteine + N1-methyladenine57/N1-methyladenine58 in tRNAAsp | - |
? | |
2 S-adenosyl-L-methionine + adenine57/adenine58 in tRNAAsp | modified mini-tRNAAsp substrate, PabTrmI recognizes and methylates both A57 and A58 in mini-tRNAAsp containing the A57A58A59U60 sequence | Pyrococcus abyssi GE5 / CNCM I-1302 / DSM 25543 | 2 S-adenosyl-L-homocysteine + N1-methyladenine57/N1-methyladenine58 in tRNAAsp | - |
? | |
additional information | methylation occurs at position 58 when position 57 contains a methylated adenine, an adenine derivative or guanine, whereas the methylation at position 57 strictly requires adenine 58 to proceed efficiently. This supports our previous conclusion that A57 is methylated before A58 in tRNAs containing the A57A58A59 sequence | Pyrococcus abyssi | ? | - |
? | |
additional information | in most organisms, the widely conserved 1-methyladenosine58 (m1A58) tRNA modification is catalyzed by S-adenosyl-L-methionine-dependent site-specific enzyme TrmI. In archaea, TrmI also methylates the adjacent adenine 57, m1A57 being an obligatory intermediate of 1-methyl-inosine57 formation, multi-site specificity mechanism, overview | Pyrococcus abyssi | ? | - |
? | |
additional information | construction of three oligoribonucleotide substrates of Pyrococcus abyssi TrmI containing a fluorescent 2-aminopurine at the two target positions 57 and 58, analysis of RNA binding kinetics and methylation reactions by stopped-flow and mass spectrometry, overview. PabTrmI does not modify 2-aminopurine but methylates the adjacent target adenine. 2-Aminopurine seriously impairs the methylation of A57 but not A58, confirming that PabTrmI methylates efficiently the first adenine of the A57A58A59 sequence | Pyrococcus abyssi | ? | - |
? | |
additional information | methylation occurs at position 58 when position 57 contains a methylated adenine, an adenine derivative or guanine, whereas the methylation at position 57 strictly requires adenine 58 to proceed efficiently. This supports our previous conclusion that A57 is methylated before A58 in tRNAs containing the A57A58A59 sequence | Pyrococcus abyssi GE5 / CNCM I-1302 / DSM 25543 | ? | - |
? | |
additional information | in most organisms, the widely conserved 1-methyladenosine58 (m1A58) tRNA modification is catalyzed by S-adenosyl-L-methionine-dependent site-specific enzyme TrmI. In archaea, TrmI also methylates the adjacent adenine 57, m1A57 being an obligatory intermediate of 1-methyl-inosine57 formation, multi-site specificity mechanism, overview | Pyrococcus abyssi GE5 / CNCM I-1302 / DSM 25543 | ? | - |
? | |
additional information | construction of three oligoribonucleotide substrates of Pyrococcus abyssi TrmI containing a fluorescent 2-aminopurine at the two target positions 57 and 58, analysis of RNA binding kinetics and methylation reactions by stopped-flow and mass spectrometry, overview. PabTrmI does not modify 2-aminopurine but methylates the adjacent target adenine. 2-Aminopurine seriously impairs the methylation of A57 but not A58, confirming that PabTrmI methylates efficiently the first adenine of the A57A58A59 sequence | Pyrococcus abyssi GE5 / CNCM I-1302 / DSM 25543 | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
bi-specific tRNA adenine-N1, A57-A58-methyltransferase | - |
Pyrococcus abyssi |
bi-specific tRNA adenine-N1, A57-A58-MTase | - |
Pyrococcus abyssi |
PAB0283 protein | - |
Pyrococcus abyssi |
PabTrmI | - |
Pyrococcus abyssi |
PYRAB04300 | locus name | Pyrococcus abyssi |
tRNA (adenine(57)-N(1)/adenine(58)-N(1))-methyltransferase | - |
Pyrococcus abyssi |
tRNA(m1A57/58)-methyltransferase | - |
Pyrococcus abyssi |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Pyrococcus abyssi |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Pyrococcus abyssi |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
S-adenosyl-L-methionine | dynamics of RNA binding by PabTrmI in the presence and absence of S-adenosyl-L-methionine | Pyrococcus abyssi |
General Information | Comment | Organism |
---|---|---|
additional information | PabTrmI methylates efficiently the first adenine of the A57A58A59 sequence. m1A58 formation triggers RNA release. A model of the protein-tRNA complex shows both target adenines in proximity of S-adenosyl-L-methionine and emphasizes no major tRNA conformational change except base flipping during the reaction. The solvent accessibility of the S-adenosyl-L-methionine pocket is not affected by the tRNA, thereby enabling S-adenosyl-L-homocysteine to be replaced by S-adenosyl-L-methionine without prior release of monomethylated tRNA. Dynamics of RNA binding by PabTrmI in the presence and absence of S-adenosyl-L-methionine, structural model of PabTrmI in complex with tRNA, overview | Pyrococcus abyssi |