Cloned (Comment) | Organism |
---|---|
gene trmD, recombinant expression of N-terminally His-tagged wild-type and S88L mutant enzymes in Escherichia coli strain SG13009 | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
S88L | naturally occuring mutation of trmD, the mutation confers thermal lability to the enzyme with a minor effect. The mutation decreases the catalytic efficiency of the enzyme to 1% of wild-type activity at permissive temperature. At nonpermissive temperature, it renders further deterioration of activity to 0.1%. These changes are accompanied by losses of both the quantity and quality of tRNA methylation, leading to the potential of cellular pleiotropic effects | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten steady-state kinetics analysis | Escherichia coli | |
0.0024 | - |
guanine37 in tRNALeu | recombinant wild-type enzyme, pH 8.0, 30°C | Escherichia coli | |
0.0031 | - |
guanine37 in tRNALeu | recombinant wild-type enzyme, pH 8.0, 37°C | Escherichia coli | |
0.013 | - |
guanine37 in tRNALeu | recombinant mutant S88L, pH 8.0, 30°C | Escherichia coli | |
0.0217 | - |
guanine37 in tRNALeu | recombinant wild-type enzyme, pH 8.0, 43°C | Escherichia coli | |
0.0522 | - |
guanine37 in tRNALeu | recombinant mutant S88L, pH 8.0, 37°C | Escherichia coli | |
0.0625 | - |
guanine37 in tRNALeu | recombinant mutant S88L, pH 8.0, 43°C | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
S-adenosyl-L-methionine + guanine37 in tRNA | Escherichia coli | - |
S-adenosyl-L-homocysteine + N1-methylguanine37 in tRNA | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P0A873 | gene trmD | - |
Purification (Comment) | Organism |
---|---|
recombinant N-terminally His-tagged wild-type and S88L mutant enzymes from Escherichia coli strain SG13009 by nickel affinity chromatography | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
S-adenosyl-L-methionine + guanine37 in tRNA | - |
Escherichia coli | S-adenosyl-L-homocysteine + N1-methylguanine37 in tRNA | - |
? | |
S-adenosyl-L-methionine + guanine37 in tRNALeu | - |
Escherichia coli | S-adenosyl-L-homocysteine + N1-methylguanine37 in tRNALeu | - |
? | |
S-adenosyl-L-methionine + guanine37 in tRNAPro | the A37 mutant of EctRNAPro is no substrate for the enzyme | Escherichia coli | S-adenosyl-L-homocysteine + N1-methylguanine37 in tRNAPro | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | TrmD exists as an obligate homodimer, with each subunit featuring an N-terminal domain (residues 1-159), a flexible linker (residues 160-169), and a C-terminal domain (residues 170-250), the active site is built between the N-terminal domain of one subunit and the flexible linker and the C-terminal domain of the other | Escherichia coli |
Synonyms | Comment | Organism |
---|---|---|
TrmD | - |
Escherichia coli |
tRNA methyltransferase D | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | 43 | assay at | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0015 | - |
guanine37 in tRNALeu | recombinant mutant S88L, pH 8.0, 43°C | Escherichia coli | |
0.0032 | - |
guanine37 in tRNALeu | recombinant mutant S88L, pH 8.0, 30°C | Escherichia coli | |
0.0067 | - |
guanine37 in tRNALeu | recombinant mutant S88L, pH 8.0, 37°C | Escherichia coli | |
0.049 | - |
guanine37 in tRNALeu | recombinant wild-type enzyme, pH 8.0, 30°C | Escherichia coli | |
0.093 | - |
guanine37 in tRNALeu | recombinant wild-type enzyme, pH 8.0, 37°C | Escherichia coli | |
0.13 | - |
guanine37 in tRNALeu | recombinant wild-type enzyme, pH 8.0, 43°C | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
S-adenosyl-L-methionine | - |
Escherichia coli |
General Information | Comment | Organism |
---|---|---|
malfunction | the ts phenotype of an essential gene mutation S88L in gene trmD can be closely linked to the catalytic defect of the gene product | Escherichia coli |
additional information | in the cross-subunit active site, S-adenosyl-L-methionine is bound to the trefoil knot fold in the N-terminal domain, whereas the target G37 is predicted to bind to the flexible linker | Escherichia coli |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.024 | - |
guanine37 in tRNALeu | recombinant mutant S88L, pH 8.0, 43°C | Escherichia coli | |
0.13 | - |
guanine37 in tRNALeu | recombinant mutant S88L, pH 8.0, 37°C | Escherichia coli | |
0.25 | - |
guanine37 in tRNALeu | recombinant mutant S88L, pH 8.0, 30°C | Escherichia coli | |
5.99 | - |
guanine37 in tRNALeu | recombinant wild-type enzyme, pH 8.0, 43°C | Escherichia coli | |
20.42 | - |
guanine37 in tRNALeu | recombinant wild-type enzyme, pH 8.0, 30°C | Escherichia coli | |
30 | - |
guanine37 in tRNALeu | recombinant wild-type enzyme, pH 8.0, 37°C | Escherichia coli |