Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Bacillus subtilis |
Protein Variants | Comment | Organism |
---|---|---|
E128A | kcat/KM for S-adenosyl-L-methionine is 46% of wild type value. kcat/KM for RNA is 59% of wild type value. No difference in generating erythromycin resistance when compared to the wild-type enzyme | Bacillus subtilis |
F163A | kcat/KM for S-adenosyl-L-methionine is 67% of wild type value. kcat/KM for RNA is 28% of wild type value. Mutant enzyme still mediates erythromycin resistance, although at reduced level | Bacillus subtilis |
K166A | kcat/KM for S-adenosyl-L-methionine is 74% of wild type value. kcat/KM for RNA is 58% of wild type value. Little difference in generating erythromycin resistance when compared to the wild-type enzyme | Bacillus subtilis |
K168A | kcat/KM for S-adenosyl-L-methionine is 59% of wild type value. kcat/KM for RNA is 70% of wild type value. No difference in generating erythromycin resistance when compared to the wild-type enzyme | Bacillus subtilis |
additional information | to validate the structure-based predictions of presumably essential residues in the catalytic pocket of ErmC', site-directed mutagenesis is carried out and the function of the mutants is studied in vitro and in vivo | Bacillus subtilis |
N101A | kcat/KM for S-adenosyl-L-methionine is 10% of wild type value. kcat/KM for RNA is 11% of wild type value. Mutant enzyme is totally unable to render DH5alpha cells resistant to erythromycin | Bacillus subtilis |
N11A | kcat/KM for S-adenosyl-L-methionine is 32% of wild type value. kcat/KM for RNA is 33% of wild type value. Little difference in generating erythromycin resistance when compared to the wild-type enzyme | Bacillus subtilis |
P165A | kcat/KM for S-adenosyl-L-methionine is 4% of wild type value. kcat/KM for RNA is 4% of wild type value. Mutant enzyme still mediates erythromycin resistance, although at reduced level | Bacillus subtilis |
Y104A | mutant enzyme is totally unable to render DH5alpha cells resistant to erythromycin | Bacillus subtilis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0003 | - |
adenine2085 in 23S rRNA | pH 7.5, 25°C, mutant enzyme E128A | Bacillus subtilis | |
0.0007 | - |
adenine2085 in 23S rRNA | pH 7.5, 25°C, wild-type enzyme | Bacillus subtilis | |
0.0008 | - |
adenine2085 in 23S rRNA | pH 7.5, 25°C, mutant enzyme N101A | Bacillus subtilis | |
0.0008 | - |
adenine2085 in 23S rRNA | pH 7.5, 25°C, mutant enzyme N11A | Bacillus subtilis | |
0.0013 | - |
adenine2085 in 23S rRNA | pH 7.5, 25°C, mutant enzyme P165A | Bacillus subtilis | |
0.0014 | - |
adenine2085 in 23S rRNA | pH 7.5, 25°C, mutant enzyme F163A | Bacillus subtilis | |
0.0014 | - |
adenine2085 in 23S rRNA | pH 7.5, 25°C, mutant enzyme K168A | Bacillus subtilis | |
0.002 | - |
S-adenosyl-L-methionine | pH 7.5, 25°C, mutant enzyme F163A | Bacillus subtilis | |
0.0028 | - |
S-adenosyl-L-methionine | pH 7.5, 25°C, wild-type enzyme | Bacillus subtilis | |
0.0031 | - |
S-adenosyl-L-methionine | pH 7.5, 25°C, mutant enzyme K166A | Bacillus subtilis | |
0.004 | - |
S-adenosyl-L-methionine | pH 7.5, 25°C, mutant enzyme K168A | Bacillus subtilis | |
0.004 | - |
S-adenosyl-L-methionine | pH 7.5, 25°C, mutant enzyme N11A | Bacillus subtilis | |
0.0041 | - |
adenine2085 in 23S rRNA | pH 7.5, 25°C, mutant enzyme K166A | Bacillus subtilis | |
0.0045 | - |
S-adenosyl-L-methionine | pH 7.5, 25°C, mutant enzyme E128A | Bacillus subtilis | |
0.0139 | - |
S-adenosyl-L-methionine | pH 7.5, 25°C, mutant enzyme N101A | Bacillus subtilis | |
0.0185 | - |
S-adenosyl-L-methionine | pH 7.5, 25°C, mutant enzyme P165A | Bacillus subtilis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus subtilis | P13956 | carrying naturally occurring plasmid pIM13 with ermC' gene | - |
Bacillus subtilis BD1167 | P13956 | carrying naturally occurring plasmid pIM13 with ermC' gene | - |
Purification (Comment) | Organism |
---|---|
recombinant enzyme | Bacillus subtilis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2 S-adenosyl-L-methionine + adenine2085 in 23S rRNA | - |
Bacillus subtilis | 2 S-adenosyl-L-homocysteine + N6-dimethyladenine2085 in 23S rRNA | - |
? | |
2 S-adenosyl-L-methionine + adenine2085 in 23S rRNA | - |
Bacillus subtilis BD1167 | 2 S-adenosyl-L-homocysteine + N6-dimethyladenine2085 in 23S rRNA | - |
? |
Synonyms | Comment | Organism |
---|---|---|
ErmC' | - |
Bacillus subtilis |
rRNA:m6A methyltransferase ErmC' | - |
Bacillus subtilis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Bacillus subtilis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.016 | - |
S-adenosyl-L-methionine | pH 7.5, 25°C, mutant enzyme P165A | Bacillus subtilis | |
0.029 | - |
S-adenosyl-L-methionine | pH 7.5, 25°C, mutant enzyme N11A | Bacillus subtilis | |
0.032 | - |
S-adenosyl-L-methionine | pH 7.5, 25°C, mutant enzyme F163A | Bacillus subtilis | |
0.033 | - |
S-adenosyl-L-methionine | pH 7.5, 25°C, mutant enzyme N101A | Bacillus subtilis | |
0.04 | - |
adenine2085 in 23S rRNA | pH 7.5, 25°C, mutant enzyme P165A | Bacillus subtilis | |
0.048 | - |
S-adenosyl-L-methionine | pH 7.5, 25°C, mutant enzyme E128A | Bacillus subtilis | |
0.054 | - |
S-adenosyl-L-methionine | pH 7.5, 25°C, mutant enzyme K166A | Bacillus subtilis | |
0.054 | - |
S-adenosyl-L-methionine | pH 7.5, 25°C, mutant enzyme K168A | Bacillus subtilis | |
0.066 | - |
S-adenosyl-L-methionine | pH 7.5, 25°C, wild-type enzyme | Bacillus subtilis | |
0.07 | - |
adenine2085 in 23S rRNA | pH 7.5, 25°C, mutant enzyme N101A | Bacillus subtilis | |
0.19 | - |
adenine2085 in 23S rRNA | pH 7.5, 25°C, mutant enzyme E128A | Bacillus subtilis | |
0.23 | - |
adenine2085 in 23S rRNA | pH 7.5, 25°C, mutant enzyme N11A | Bacillus subtilis | |
0.37 | - |
adenine2085 in 23S rRNA | pH 7.5, 25°C, mutant enzyme F163A | Bacillus subtilis | |
0.64 | - |
adenine2085 in 23S rRNA | pH 7.5, 25°C, wild-type enzyme | Bacillus subtilis | |
0.87 | - |
adenine2085 in 23S rRNA | pH 7.5, 25°C, mutant enzyme K168A | Bacillus subtilis | |
2.2 | - |
adenine2085 in 23S rRNA | pH 7.5, 25°C, mutant enzyme K166A | Bacillus subtilis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Bacillus subtilis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.86 | - |
S-adenosyl-L-methionine | pH 7.5, 25°C, mutant enzyme P165A | Bacillus subtilis | |
2.4 | - |
S-adenosyl-L-methionine | pH 7.5, 25°C, mutant enzyme N101A | Bacillus subtilis | |
7.3 | - |
S-adenosyl-L-methionine | pH 7.5, 25°C, mutant enzyme N11A | Bacillus subtilis | |
10.7 | - |
S-adenosyl-L-methionine | pH 7.5, 25°C, mutant enzyme E128A | Bacillus subtilis | |
13.5 | - |
S-adenosyl-L-methionine | pH 7.5, 25°C, mutant enzyme K168A | Bacillus subtilis | |
16 | - |
S-adenosyl-L-methionine | pH 7.5, 25°C, mutant enzyme F163A | Bacillus subtilis | |
17.4 | - |
S-adenosyl-L-methionine | pH 7.5, 25°C, mutant enzyme K166A | Bacillus subtilis | |
23.6 | - |
S-adenosyl-L-methionine | pH 7.5, 25°C, wild-type enzyme | Bacillus subtilis | |
30.7 | - |
adenine2085 in 23S rRNA | pH 7.5, 25°C, mutant enzyme P165A | Bacillus subtilis | |
87.5 | - |
adenine2085 in 23S rRNA | pH 7.5, 25°C, mutant enzyme N101A | Bacillus subtilis | |
264 | - |
adenine2085 in 23S rRNA | pH 7.5, 25°C, mutant enzyme F163A | Bacillus subtilis | |
287.5 | - |
adenine2085 in 23S rRNA | pH 7.5, 25°C, mutant enzyme N11A | Bacillus subtilis | |
536 | - |
adenine2085 in 23S rRNA | pH 7.5, 25°C, mutant enzyme K166A | Bacillus subtilis | |
621 | - |
adenine2085 in 23S rRNA | pH 7.5, 25°C, mutant enzyme K168A | Bacillus subtilis | |
633 | - |
adenine2085 in 23S rRNA | pH 7.5, 25°C, mutant enzyme E128A | Bacillus subtilis | |
914 | - |
adenine2085 in 23S rRNA | pH 7.5, 25°C, wild-type enzyme | Bacillus subtilis |