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Literature summary for 2.1.1.184 extracted from

  • Maravic, G.; Feder, M.; Pongor, S.; Flogel, M.; Bujnicki, J.M.
    Mutational analysis defines the roles of conserved amino acid residues in the predicted catalytic pocket of the rRNA:m6A methyltransferase ErmC' (2003), J. Mol. Biol., 332, 99-109.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Bacillus subtilis

Protein Variants

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 [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

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 (Commentary)

Purification (Comment) Organism
recombinant enzyme Bacillus subtilis

Substrates and Products (Substrate)

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

Synonyms Comment Organism
ErmC'
-
Bacillus subtilis
rRNA:m6A methyltransferase ErmC'
-
Bacillus subtilis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Bacillus subtilis

Turnover Number [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at Bacillus subtilis

kcat/KM [mM/s]

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