Protein Variants | Comment | Organism |
---|---|---|
D223A | kcat/Km for guanine37 in Methanocaldococcus jannaschii tRNACys is 1% of the wild-type value | Methanocaldococcus jannaschii |
G205A/G207A | kcat/Km for guanine37 in Methanocaldococcus jannaschii tRNACys is 3% of the wild-type value | Methanocaldococcus jannaschii |
additional information | proline at position 267 is a critical residue for catalysis, because substitution of this residue severely decreases kcat of the methylation reaction in steady-state kinetic analysis. However, substitution of P267 has milder effect on Km and little effect on Kd of either substrate. Because P267 has no functional side chain that can directly participate in the chemistry of methyl transfer, we suggest that its role in catalysis is to stabilize conformations of enzyme and substrates for proper alignment of reactive groups at the enzyme active site. Sequence analysis shows that P267 is embedded in a peptide motif that is conserved among the Trm5 family, but absent from the TrmD family, supporting the notion that the two families are descendants of unrelated protein structures | Methanocaldococcus jannaschii |
N225A | kcat/Km for guanine37 in Methanocaldococcus jannaschii tRNACys is 5% of the wild-type value | Methanocaldococcus jannaschii |
N265A | kcat/Km for guanine37 in Methanocaldococcus jannaschii tRNACys is 1% of the wild-type value | Methanocaldococcus jannaschii |
P226A | kcat/Km for guanine37 in Methanocaldococcus jannaschii tRNACys is 6% of the wild-type value | Methanocaldococcus jannaschii |
P267A | kcat/Km for guanine37 in Methanocaldococcus jannaschii tRNACys is 0.1% of the wild-type value | Methanocaldococcus jannaschii |
R144A | kcat/Km for guanine37 in Methanocaldococcus jannaschii tRNACys is 6% of the wild-type value | Methanocaldococcus jannaschii |
Y176A | kcat/Km for guanine37 in Methanocaldococcus jannaschii tRNACys is 5% of the wild-type value | Methanocaldococcus jannaschii |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0005 | - |
S-adenosyl-L-methionine | pH 8.0, 50°C, mutant enzyme P267A | Methanocaldococcus jannaschii | |
0.0007 | - |
guanine37 in Methanocaldococcus jannaschii tRNACys | pH 8.0, 50°C, wild-type enzyme | Methanocaldococcus jannaschii | |
0.001 | - |
S-adenosyl-L-methionine | pH 8.0, 50°C, wild-type enzyme | Methanocaldococcus jannaschii | |
0.0025 | - |
guanine37 in Methanocaldococcus jannaschii tRNACys | pH 8.0, 50°C, mutant enzyme Y176A | Methanocaldococcus jannaschii | |
0.003 | - |
guanine37 in Methanocaldococcus jannaschii tRNACys | pH 8.0, 50°C, mutant enzyme D223A | Methanocaldococcus jannaschii | |
0.003 | - |
guanine37 in Methanocaldococcus jannaschii tRNACys | pH 8.0, 50°C, mutant enzyme P226A | Methanocaldococcus jannaschii | |
0.0043 | - |
guanine37 in Methanocaldococcus jannaschii tRNACys | pH 8.0, 50°C, mutant enzyme R144A | Methanocaldococcus jannaschii | |
0.0046 | - |
guanine37 in Methanocaldococcus jannaschii tRNACys | pH 8.0, 50°C, mutant enzyme G205A/G207A | Methanocaldococcus jannaschii | |
0.006 | - |
guanine37 in Methanocaldococcus jannaschii tRNACys | pH 8.0, 50°C, mutant enzyme P267A | Methanocaldococcus jannaschii | |
0.0072 | - |
guanine37 in Methanocaldococcus jannaschii tRNACys | pH 8.0, 50°C, mutant enzyme N225A | Methanocaldococcus jannaschii | |
0.0083 | - |
guanine37 in Methanocaldococcus jannaschii tRNACys | pH 8.0, 50°C, mutant enzyme N265A | Methanocaldococcus jannaschii |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Methanocaldococcus jannaschii | Q58293 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Methanocaldococcus jannaschii |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
S-adenosyl-L-methionine + guanine37 in Methanocaldococcus jannaschii tRNACys | - |
Methanocaldococcus jannaschii | S-adenosyl-L-homocysteine + N1-methylguanine37 in Methanocaldococcus jannaschii tRNACys | - |
? |
Synonyms | Comment | Organism |
---|---|---|
TRM5 | - |
Methanocaldococcus jannaschii |
tRNA(m1G37) methyltransferase | - |
Methanocaldococcus jannaschii |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | - |
assay | Methanocaldococcus jannaschii |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0001 | - |
S-adenosyl-L-methionine | pH 8.0, 50°C, mutant enzyme P267A | Methanocaldococcus jannaschii | |
0.0001 | - |
guanine37 in Methanocaldococcus jannaschii tRNACys | pH 8.0, 50°C, mutant enzyme P267A | Methanocaldococcus jannaschii | |
0.0003 | - |
guanine37 in Methanocaldococcus jannaschii tRNACys | pH 8.0, 50°C, mutant enzyme D223A | Methanocaldococcus jannaschii | |
0.0013 | - |
guanine37 in Methanocaldococcus jannaschii tRNACys | pH 8.0, 50°C, mutant enzyme N265A | Methanocaldococcus jannaschii | |
0.0013 | - |
guanine37 in Methanocaldococcus jannaschii tRNACys | pH 8.0, 50°C, mutant enzyme Y176A | Methanocaldococcus jannaschii | |
0.0015 | - |
guanine37 in Methanocaldococcus jannaschii tRNACys | pH 8.0, 50°C, mutant enzyme G205A/G207A | Methanocaldococcus jannaschii | |
0.0018 | - |
guanine37 in Methanocaldococcus jannaschii tRNACys | pH 8.0, 50°C, mutant enzyme P226A | Methanocaldococcus jannaschii | |
0.003 | - |
guanine37 in Methanocaldococcus jannaschii tRNACys | pH 8.0, 50°C, mutant enzyme R144A | Methanocaldococcus jannaschii | |
0.0037 | - |
guanine37 in Methanocaldococcus jannaschii tRNACys | pH 8.0, 50°C, mutant enzyme N225A | Methanocaldococcus jannaschii | |
0.0083 | - |
guanine37 in Methanocaldococcus jannaschii tRNACys | pH 8.0, 50°C, wild-type enzyme | Methanocaldococcus jannaschii | |
0.012 | - |
S-adenosyl-L-methionine | pH 8.0, 50°C, wild-type enzyme | Methanocaldococcus jannaschii |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Methanocaldococcus jannaschii |
General Information | Comment | Organism |
---|---|---|
physiological function | modification at guanine37 is important for maintaining the reading frame fidelity | Methanocaldococcus jannaschii |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.017 | - |
guanine37 in Methanocaldococcus jannaschii tRNACys | pH 8.0, 50°C, mutant enzyme P267A | Methanocaldococcus jannaschii | |
0.11 | - |
guanine37 in Methanocaldococcus jannaschii tRNACys | pH 8.0, 50°C, mutant enzyme D223A | Methanocaldococcus jannaschii | |
0.16 | - |
guanine37 in Methanocaldococcus jannaschii tRNACys | pH 8.0, 50°C, mutant enzyme N265A | Methanocaldococcus jannaschii | |
0.2 | - |
S-adenosyl-L-methionine | pH 8.0, 50°C, mutant enzyme P267A | Methanocaldococcus jannaschii | |
0.32 | - |
guanine37 in Methanocaldococcus jannaschii tRNACys | pH 8.0, 50°C, mutant enzyme G205A/G207A | Methanocaldococcus jannaschii | |
0.51 | - |
guanine37 in Methanocaldococcus jannaschii tRNACys | pH 8.0, 50°C, mutant enzyme N225A | Methanocaldococcus jannaschii | |
0.53 | - |
guanine37 in Methanocaldococcus jannaschii tRNACys | pH 8.0, 50°C, mutant enzyme Y176A | Methanocaldococcus jannaschii | |
0.61 | - |
guanine37 in Methanocaldococcus jannaschii tRNACys | pH 8.0, 50°C, mutant enzyme P226A | Methanocaldococcus jannaschii | |
0.775 | - |
guanine37 in Methanocaldococcus jannaschii tRNACys | pH 8.0, 50°C, mutant enzyme R144A | Methanocaldococcus jannaschii | |
11.9 | - |
guanine37 in Methanocaldococcus jannaschii tRNACys | pH 8.0, 50°C, wild-type enzyme | Methanocaldococcus jannaschii | |
12.2 | - |
S-adenosyl-L-methionine | pH 8.0, 50°C, wild-type enzyme | Methanocaldococcus jannaschii |