Any feedback?
Information on EC 2.1.1.184 - 23S rRNA (adenine2085-N6)-dimethyltransferase and Organism(s) Bacillus subtilis and UniProt Accession P13956
for references in articles please use BRENDA:EC2.1.1.184Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
EC Tree
2.1.1.184 23S rRNA (adenine2085-N6)-dimethyltransferaseIUBMB Comments
ErmC is a methyltransferase that confers resistance to the macrolide-lincosamide-streptogramin B group of antibiotics by catalysing the methylation of 23S rRNA at adenine2085.
Specify your search results
Word Map
- 2.1.1.184
-
macrolide
-
macrolide-lincosamide-streptogramin
- methyltransferases
-
lincosamide
-
unmethylated
- mtases
-
streptogramine
- medicine
The taxonomic range for the selected organisms is: Bacillus subtilis
The expected taxonomic range for this enzyme is: Bacteria, Archaea
The expected taxonomic range for this enzyme is: Bacteria, Archaea
Reaction Schemes
2
+
=
2
+
2
+
adenine2085 in 23S rRNA
=
2
+
N6-dimethyladenine2085 in 23S rRNA
Synonyms
ermc', ermc methylase, ermc' methyltransferase, ermc methyltransferase, erythromycin resistance protein, rrna methyltransferase ermc', ermc 23s rrna methyltransferase, ermc 23 s rrna methyltransferase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
ErmC'
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
2 S-adenosyl-L-methionine + adenine2085 in 23S rRNA = 2 S-adenosyl-L-homocysteine + N6-dimethyladenine2085 in 23S rRNA
2 S-adenosyl-L-methionine + adenine2085 in 23S rRNA = 2 S-adenosyl-L-homocysteine + N6-dimethyladenine2085 in 23S rRNA
a model for the transition-state based on the atomic details of the active site is consistent with a one-step methyl-transfer
2 S-adenosyl-L-methionine + adenine2085 in 23S rRNA = 2 S-adenosyl-L-homocysteine + N6-dimethyladenine2085 in 23S rRNA
a model of RNA-ErmC interaction involving multiple binding sites is proposed from the kinetic data
2 S-adenosyl-L-methionine + adenine2085 in 23S rRNA = 2 S-adenosyl-L-homocysteine + N6-dimethyladenine2085 in 23S rRNA
ermC methylase reaction involves a sequential mechanism occurring by two consecutive random bi bi reactions
SYSTEMATIC NAME
IUBMB Comments
S-adenosyl-L-methionine:23S rRNA (adenine2085-N6)-dimethyltransferase
ErmC is a methyltransferase that confers resistance to the macrolide-lincosamide-streptogramin B group of antibiotics by catalysing the methylation of 23S rRNA at adenine2085.
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
2 S-adenosyl-L-methionine + adenine2085 in 23S rRNA
2 S-adenosyl-L-homocysteine + N6-dimethyladenine2085 in 23S rRNA
2 S-adenosyl-L-methionine + adenine2085 in a 262-nucleotide RNA fragment within domain V of Bacillus subtilis 23S rRNA
2 S-adenosyl-L-homocysteine + N6-dimethyladenine2085 in 262-nucleotide RNA fragment within domain V of Bacillus subtilis 23S rRNA
a 262-nucleotide RNA fragment within domain V of Bacillus subtilis 23S rRNA can be utilized efficiently as a substrate for methylation at adenine2085
-
-
?
2 S-adenosyl-L-methionine + adenine2085 in a 623-nucleotide RNA fragment within domain V of Bacillus subtilis 23S rRNA
2 S-adenosyl-L-homocysteine + N6-dimethyladenine2085 in a 623-nucleotide RNA fragment within domain V of Bacillus subtilis 23S rRNA
-
-
-
?
2 S-adenosyl-L-methionine + adenine2085 in a 623-nucleotide RNA fragment within domain V of Bacillus subtilis 23S rRNA with mutation A2086T
2 S-adenosyl-L-homocysteine + N6-dimethyladenine2085 in a 623-nucleotide RNA fragment within domain V of Bacillus subtilis 23S rRNA with mutation A2086T
the A2086T change is methylated to ca. 50% of the level of wild-type domain V
-
-
?
S-adenosyl-L-methionine + 23S rRNA
S-adenosyl-L-homocysteine + 23S rRNA containing N6-dimethyladenine
-
-
-
?
S-adenosyl-L-methionine + 262-nucleotide RNA fragment within domain V of Bacillus subtilis 23S rRNA
S-adenosyl-L-homocysteine + 262-nucleotide RNA fragment within domain V of Bacillus subtilis 23S rRNA containing N6-dimethyladenine
-
-
-
?
S-adenosyl-L-methionine + 623-nucleotide RNA fragment within domain V of Bacillus subtilis 23S rRNA
S-adenosyl-L-homocysteine + 623-nucleotide RNA fragment within domain V of Bacillus subtilis 23S rRNA containing N6-dimethyladenine
-
-
-
?
S-adenosyl-L-methionine + 623-nucleotide RNA fragment within domain V of Bacillus subtilis 23S rRNA with mutation A2086T
S-adenosyl-L-homocysteine + 623-nucleotide RNA fragment within domain V of Bacillus subtilis 23S rRNA with mutation A2086T containing N6-dimethyladenine
-
-
-
?
2 S-adenosyl-L-methionine + adenine2085 in 23S rRNA
2 S-adenosyl-L-homocysteine + N6-dimethyladenine2085 in 23S rRNA
-
-
-
?
2 S-adenosyl-L-methionine + adenine2085 in 23S rRNA
2 S-adenosyl-L-homocysteine + N6-dimethyladenine2085 in 23S rRNA
-
-
-
-
?
2 S-adenosyl-L-methionine + adenine2085 in 23S rRNA
2 S-adenosyl-L-homocysteine + N6-dimethyladenine2085 in 23S rRNA
-
-
-
?
2 S-adenosyl-L-methionine + adenine2085 in 23S rRNA
2 S-adenosyl-L-homocysteine + N6-dimethyladenine2085 in 23S rRNA
ErmC is a methyltransferase that confers resistance to the macrolide-lincosamide-streptogramin B group of antibiotics by catalyzing the methylation of 23S rRNA at a specific adenine residue (A2085 in Bacillus subtilis)
-
-
?
2 S-adenosyl-L-methionine + adenine2085 in 23S rRNA
2 S-adenosyl-L-homocysteine + N6-dimethyladenine2085 in 23S rRNA
the rRNA methyltransferase ErmC0 transfers methyl groups from S-adenosyl-L-methionine to atom N6 of an adenine base within the peptidyltransferase loop of 23 S rRNA, thus conferring antibiotic resistance against a number of macrolide antibiotics
-
-
?
2 S-adenosyl-L-methionine + adenine2085 in 23S rRNA
2 S-adenosyl-L-homocysteine + N6-dimethyladenine2085 in 23S rRNA
ermC methyltransferase produces both N6-mono and N6,N6-dimethylated adenine residues in Bacillus subtilis 23 S rRNA during the course of the reaction in vitro. The addition of the two methyl groups to each 23 S rRNA molecule takes place through a monomethylated intermediate and suggest that the enzyme dissociates from its RNA substrate between the two consecutive methylation reactions. The enzyme is able to utilize monomethylated RNA as substrate for the addition of a second methyl group with an efficiency approximately comparable to that obtained when unmethylated RNA is the initial substrate
-
-
?
2 S-adenosyl-L-methionine + adenine2085 in 23S rRNA
2 S-adenosyl-L-homocysteine + N6-dimethyladenine2085 in 23S rRNA
since methyl groups are incorporated in protein-free 23S rRNA molecules, the structure of rRNA alone must contain sufficient information to specify the methylation site. Highest incorporation is obtained with Bacillus subtilis 23S rRNA. Escherichia coli 23S rRNA acts as a poorer substrate (35% of the methylation obtained with Bacillus subtilis 23S rRNA)
-
-
?
2 S-adenosyl-L-methionine + adenine2085 in 23S rRNA
2 S-adenosyl-L-homocysteine + N6-dimethyladenine2085 in 23S rRNA
synthetic 32-nt RNA oligonucleotide (5-GCGACGGACGGA2085AAGACCCCUAUCCGUCGCG-3, hairpin structure) designed to mimic the adenine loop in domain V of Bacillus subtilis 23S rRNA (residues 20732090 and 26382651)
-
-
?
2 S-adenosyl-L-methionine + adenine2085 in 23S rRNA
2 S-adenosyl-L-homocysteine + N6-dimethyladenine2085 in 23S rRNA
the specificity of methylation on adenine2085 is confirmed by site-directed mutagenesis. All three mutated domain V fragments, A2085T, A2085G, and A2085C, are methylated to less than 10% of the level observed with the correct domain V RNA fragment. The G2084A change reduces the methylation of the resultant domain V fragment to ca. 12% of the level of the wild-type domain V fragment
-
-
?
additional information
?
-
direct autoregulatory mechanism operating at the posttranscriptional level and independently of the ermC methylase-mediated methylation of ribosomes. A translational repression model is suggested in which the ermC methyltransferase binds to its own mRNA, at a region that resembles the methylation target site on 23S rRNA
-
-
?
additional information
?
-
-
direct autoregulatory mechanism operating at the posttranscriptional level and independently of the ermC methylase-mediated methylation of ribosomes. A translational repression model is suggested in which the ermC methyltransferase binds to its own mRNA, at a region that resembles the methylation target site on 23S rRNA
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
2 S-adenosyl-L-methionine + adenine2085 in 23S rRNA
2 S-adenosyl-L-homocysteine + N6-dimethyladenine2085 in 23S rRNA
2 S-adenosyl-L-methionine + adenine2085 in 23S rRNA
2 S-adenosyl-L-homocysteine + N6-dimethyladenine2085 in 23S rRNA
-
-
-
?
2 S-adenosyl-L-methionine + adenine2085 in 23S rRNA
2 S-adenosyl-L-homocysteine + N6-dimethyladenine2085 in 23S rRNA
ErmC is a methyltransferase that confers resistance to the macrolide-lincosamide-streptogramin B group of antibiotics by catalyzing the methylation of 23S rRNA at a specific adenine residue (A2085 in Bacillus subtilis)
-
-
?
2 S-adenosyl-L-methionine + adenine2085 in 23S rRNA
2 S-adenosyl-L-homocysteine + N6-dimethyladenine2085 in 23S rRNA
the rRNA methyltransferase ErmC0 transfers methyl groups from S-adenosyl-L-methionine to atom N6 of an adenine base within the peptidyltransferase loop of 23 S rRNA, thus conferring antibiotic resistance against a number of macrolide antibiotics
-
-
?
additional information
?
-
direct autoregulatory mechanism operating at the posttranscriptional level and independently of the ermC methylase-mediated methylation of ribosomes. A translational repression model is suggested in which the ermC methyltransferase binds to its own mRNA, at a region that resembles the methylation target site on 23S rRNA
-
-
?
additional information
?
-
-
direct autoregulatory mechanism operating at the posttranscriptional level and independently of the ermC methylase-mediated methylation of ribosomes. A translational repression model is suggested in which the ermC methyltransferase binds to its own mRNA, at a region that resembles the methylation target site on 23S rRNA
-
-
?
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
2-([[(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)methyl][3-(1H-imidazol-1-yl)propyl]amino]methyl)-1H-isoindole-1,3(2H)-dione
i.e. PD00556
N6-dimethyladenine2085 containing 23S rRNA
linear competitive inhibition kinetics with RNA as the variable substrate, mixed inhibition with S-adenosyl-L-methionine as the variable substrate
-
nicotinaldehyde-N-[3-(2-chlorobenzyl)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-7-yl]hydrazone
i.e. HTS12610
additional information
the crystal structure of ErmC methyltransferase is used as a target for structure-based virtual screening of a database composed of 58679 lead-like compounds. Among 77 compounds selected for experimental validation (63 predicted to bind to the catalytic pocket and 14 compounds predicted to bind to the putative RNA binding site), several novel inhibitors are found that decrease the minimal inhibitory concentration of a macrolide antibiotic erythromycin toward an Escherichia coli strain that constitutively expresses ErmC'. Analysis of docking models of the identified inhibitors suggests a novel strategy to develop potent and clinically useful inhibitors
-
S-adenosyl-L-homocysteine
linear competitive pattern with S-adenosyl-L-methionine as the variable substrate, and a mixed inhibition kinetics with RNA
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0000375
23S rRNA
pH 7.5, 37°C, a 262-nucleotide RNA fragment within domain V of Bacillus subtilis 23S rRNA
-
0.00091
262-nucleotide RNA fragment within domain V of Bacillus subtilis 23S rRNA
pH 7.5, 37°C, a 262-nucleotide RNA fragment within domain V of Bacillus subtilis 23S rRNA
-
0.0000344
623-nucleotide RNA fragment within domain V of Bacillus subtilis 23S rRNA
pH 7.5, 37°C, a 262-nucleotide RNA fragment within domain V of Bacillus subtilis 23S rRNA
-
0.000144
623-nucleotide RNA fragment within domain V of Bacillus subtilis 23S rRNA with mutation A2086T
pH 7.5, 37°C, a 262-nucleotide RNA fragment within domain V of Bacillus subtilis 23S rRNA
-
0.00121
adenine2085 in 23S rRNA
pH 7.5, 25°C, mutant enzyme K197A/N200A/E202A/K204A/K205A
-
0.0028
S-adenosyl-L-methionine
pH 7.5, 25°C, wild-type enzyme
0.0066
S-adenosyl-L-methionine
pH 7.5, 25°C, mutant enzyme K197A/N200A/E202A/K204A/K205A
additional information
additional information
interaction between 23s rRNA and ermC' methyltransferase. Kinetic and thermodynamic parameters of binding are determined
-
additional information
additional information
-
interaction between 23s rRNA and ermC' methyltransferase. Kinetic and thermodynamic parameters of binding are determined
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.00193
adenine2085 in 23S rRNA
pH 7.5, 25°C, mutant enzyme K197A/N200A/E202A/K204A/K205A
-
0.0314
S-adenosyl-L-methionine
pH 7.5, 25°C, mutant enzyme K197A/N200A/E202A/K204A/K205A
additional information
additional information
interaction between 23s rRNA and ermC' methyltransferase. Kinetic and thermodynamic parameters of binding are determined
-
additional information
additional information
-
interaction between 23s rRNA and ermC' methyltransferase. Kinetic and thermodynamic parameters of binding are determined
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
1.6
adenine2085 in 23S rRNA
pH 7.5, 25°C, mutant enzyme K197A/N200A/E202A/K204A/K205A
-
4.79
S-adenosyl-L-methionine
pH 7.5, 25°C, mutant enzyme K197A/N200A/E202A/K204A/K205A
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.000128
N6-dimethyladenine2085 containing 23S rRNA
pH 7.5, 37°C, S-adenosyl-L-methionine as the variable substrate
-
0.00036
N6-dimethyladenine2085 containing 23S rRNA
pH 7.5, 37°C, with RNA as the variable substrate
-
0.069
S-adenosyl-L-homocysteine
pH 7.5, 37°C, with RNA as the variable substrate
0.086
S-adenosyl-L-homocysteine
pH 7.5, 37°C, S-adenosyl-L-methionine as the variable substrate
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.3
2-([[(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)methyl][3-(1H-imidazol-1-yl)propyl]amino]methyl)-1H-isoindole-1,3(2H)-dione
Bacillus subtilis
pH 7.5, 25°C
0.18
4-methyl-2,6-di[(4-methylphenyl)thio]nicotinonitrile
Bacillus subtilis
pH 7.5, 25°C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7.5
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
37
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
carrying naturally occurring plasmid pIM13 with ermC' gene
SwissProt
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
physiological function
physiological function
ErmC is a methyltransferase that confers resistance to the macrolide-lincosamide-streptogramin B group of antibiotics by catalyzing the methylation of 23S rRNA at a specific adenine residue (A2085 in Bacillus subtilis)
physiological function
the enzyme confers resistance to macrolide-lincosamide-streptogramin B antibiotics
physiological function
transfers methyl groups from S-adenosyl-l-methionine to atom N6 of an adenine base within the peptidyltransferase loop of 23S rRNA, thus conferring antibiotic resistance against a number of macrolide antibiotics
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
crystallized by the hanging drop vapor diffusion method. Structure of the apo-enzyme at 2.2 A resolution. The crystal structures of ErmC' and of its complexes with the cofactor S-adenosyl-L-methionine, the reaction product S-adenosyl-L-homocysteine and the methyltransferase inhibitor sinefungin, respectively, show that the enzyme undergoes small conformational changes upon ligand binding
crystals of ErmC' are obtained by the hanging-drop vapor diffusion method. Crystal structure of ErmC' (a naturally occurring variant of ErmC) determined at 3.0 A resolution by multiple anomalous diffraction phasing methods. The structure consists of a conserved alpha/beta amino-terminal domain which binds the cofactor S-adenosyl-L-methionine, followed by a smaller, alpha-helical RNA-recognition domain
the crystal structure of ErmC methyltransferase is used as a target for structure-based virtual screening of a database composed of 58679 lead-like compounds. Analysis of docking models of the identified inhibitors suggests a novel strategy to develop potent and clinically useful inhibitors
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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
E202A
mutant of a residues positioned on the surface of the small domain, does not display substantial defects in activity compared with the wild-type enzyme. kcat/Km for S-adenosyl-L-methionine is 32% of the wild-type value. kcat/Km for RNA is 40% of wild-type value
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
K133A
decreased ability in rendering DH5alpha cells resistant to erythromycin, suggesting that this amino acid is not absolutely indispensable for the ErmC' activity, but might be involved in important RNA-protein interactions. No change in affinity towards the RNA substrate. kcat/Km for S-adenosyl-L-methionine is% of the wild-type value. kcat/Km for RNA is 27% of wild-type value. kcat/Km for S-adenosyl-L-methionine is 13% of the wild-type value. kcat/Km for RNA is% of wild-type value
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
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
K197A
mutant of a residues positioned on the surface of the small domain, does not display substantial defects in activity compared with the wild-type enzyme. kcat/Km for S-adenosyl-L-methionine is 67% of the wild-type value. kcat/Km for RNA is 50% of wild-type value
K197A/N200A/E202A/K204A/K205A
the five mutations together do not show a visible cumulative effect on the ErmC' activity in vivo
K204A
mutant of a residues positioned on the surface of the small domain, does not display substantial defects in activity compared with the wild-type enzyme. kcat/Km for S-adenosyl-L-methionine is 73% of the wild-type value. kcat/Km for RNA is 89% of wild-type value
K205A
mutant of a residues positioned on the surface of the small domain, does not display substantial defects in activity compared with the wild-type enzyme. kcat/Km for S-adenosyl-L-methionine is 63% of the wild-type value. kcat/Km for RNA is 92% of wild-type value
K209A
mutant of a residues positioned on the surface of the small domain, does not display substantial defects in activity compared with the wild-type enzyme. kcat/Km for S-adenosyl-L-methionine is 72% of the wild-type value. kcat/Km for RNA is 73% of wild-type value
M196A
mutant of a residues positioned on the surface of the small domain, does not display substantial defects in activity compared with the wild-type enzyme. kcat/Km for S-adenosyl-L-methionine is 42% of the wild-type value. kcat/Km for RNA is 56% of wild-type value
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
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
N192A
mutant of a residues positioned on the surface of the small domain, does not display substantial defects in activity compared with the wild-type enzyme. kcat/Km for S-adenosyl-L-methionine is 73% of the wild-type value. kcat/Km for RNA is 74% of wild-type value
N200A
mutant of a residues positioned on the surface of the small domain, does not display substantial defects in activity compared with the wild-type enzyme. kcat/Km for S-adenosyl-L-methionine is 42% of the wild-type value. kcat/Km for RNA is 54% of wild-type value
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
R112A
decreased ability in rendering DH5alpha cells resistant to erythromycin, suggesting that this amino acid is not absolutely indispensable for the ErmC' activity, but might be involved in important RNA-protein interactions. kcat/Km for S-adenosyl-L-methionine is 12% of the wild-type value. kcat/Km for RNA is 7% of wild-type value
R112D
decreased ability in rendering DH5alpha cells resistant to erythromycin. The R112D mutant shows a more pronounced decrease in RNA-binding affinity compared with R112A. kcat/Km for S-adenosyl-L-methionine is 5% of the wild-type value. kcat/Km for RNA is 2% of wild-type value
R134A
mutant exhibits the most severe effect on the ErmC' ability to generate erythromycin resistance, this mutant has completely lost the activity in vivo. kcat/Km for S-adenosyl-L-methionine is 3% of the wild-type value. kcat/Km for RNA is 1% of wild-type value
R140A
decreased ability in rendering DH5alpha cells resistant to erythromycin, suggesting that this amino acid is not absolutely indispensable for the ErmC' activity, but might be involved in important RNA-protein interactions. kcat/Km for S-adenosyl-L-methionine is 1% of the wild-type value. kcat/Km for RNA is 3% of wild-type value
T108A
decreased ability in rendering DH5alpha cells resistant to erythromycin, suggesting that this amino acid is not absolutely indispensable for the ErmC' activity, but might be involved in important RNA-protein interactions. kcat/Km for S-adenosyl-L-methionine is 4% of the wild-type value. kcat/Km for RNA is 3% of wild-type value
Y104A
mutant enzyme is totally unable to render DH5alpha cells resistant to erythromycin
additional information
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
additional information
With the aim of identification of essential protein-RNA interactions, charged side chains on the predicted target-binding surface of ErmC' are replaced systematically with alanine and the function of the single- and multiple-site mutants is studied in vitro and in vivo. kcat/Km for S-adenosyl-L-methionine is 21% of the wild-type value. kcat/Km for RNA is 17% of wild-type value
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant enzyme
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
the expression of ErmC' by translational coupling to kdsB, under the control of the T7lac promoter. The pTERM31 plasmid is transformed into Escherichia coli strain BL219(DE3)/pLysS
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
direct autoregulatory mechanism operating at the posttranscriptional level and independently of the ermC methylase-mediated methylation of ribosomes. A translational repression model is suggested in which the ermC methyltransferase binds to its own mRNA, at a region that resembles the methylation target site on 23S rRNA
induction is due to a posttranscriptional mechanism in which the inducer activates translation of methylase mRNA by binding to unmethylated (erythromycin-sensitive) ribosomes and stalling them in the leader region. Pseudomonic acid A can also induce methylase synthesis. Isoleucine starvation has a similar effect on ribosomes translating the ermC leader region to cause induction of methylase synthesis. Requirement for ribosome stalling and destabilization of a stem-loop structure and demonstration that stalling can occur without macrolide-bound ribosomes
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
medicine
medicine
ErmC' catalyzes S-adenosyl-L-methionine-dependent modification of a specific adenine residue in bacterial 23S rRNA, thereby conferring resistance to clinically important macrolide, lincosamide, and streptogramin B antibiotics. The crystal structure of ErmC methyltransferase is used as a target for structure-based virtual screening of a database composed of 58679 lead-like compounds. Among 77 compounds selected for experimental validation (63 predicted to bind to the catalytic pocket and 14 compounds predicted to bind to the putative RNA bindingsite), several novel inhibitors are found that decrease the minimal inhibitory concentration of a macrolide antibiotic erythromycin toward an Escherichia coli strain that constitutively expresses ErmC'. Analysis of docking models of the identified inhibitors suggests a novel strategy to develop potent and clinically useful inhibitors
medicine
the Erm family of adenine-N6 methyltransferases is responsible for the development of resistance to macrolide-lincosamide-streptogramin B antibiotics through the methylation of 23S ribosomal RNA. Hence, these proteins are important potential drug targets
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Denoya, C.; Dubnau, D.
Site and substrate specificity of the ermC 23S rRNA methyltransferase
J. Bacteriol.
169
3857-3860
1987
Bacillus subtilis (P13956), Bacillus subtilis
Denoya, C.; Dubnau, D.
Mono- and dimethylating activities and kinetic studies of the ermC 23 S rRNA methyltransferase
J. Biol. Chem.
264
2615-2624
1989
Bacillus subtilis (P13956), Bacillus subtilis BD170 (P13956)
Bussiere, D.E.; Muchmore, S.W.; Dealwis, C.G.; Schluckebier, G.; Nienaber, V.L.; Edalji, R.P.; Walter, K.A.; Ladror, U.S.; Holzman, T.F.; Abad-Zapatero, C.
Crystal structure of ErmC', an rRNA methyltransferase which mediates antibiotic resistance in bacteria
Biochemistry
37
7103-7112
1998
Bacillus subtilis (P13956), Bacillus subtilis
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'
J. Mol. Biol.
332
99-109
2003
Bacillus subtilis (P13956), Bacillus subtilis BD1167 (P13956)
Su, S.L.
Dubnau, D.: Binding of Bacillus subtilis ermC' methyltransferase to 23S rRNA
Biochemistry
29
6033-6042
1990
Bacillus subtilis (P13956), Bacillus subtilis
Feder, M.; Purta, E.; Koscinski, L.; Ebrilo, S.; Vlahovicek, G.; Bujnicki, J.
Virtual screening and experimental verification to identify potential inhibitors of the ErmC methyltransferase responsible for bacterial resistance against macrolide antibiotics
ChemMedChem
3
316-322
2008
Bacillus subtilis (P13956)
Denoya, C.D.; Bechhofer, D.H.; Dubnau, D.
Translational autoregulation of ermC 23S rRNA methyltransferase expression in Bacillus subtilis
J. Bacteriol.
168
1133-1141
1986
Bacillus subtilis (P13956), Bacillus subtilis
Kadam, S.K.
Induction of ermC methylase in the absence of macrolide antibiotics and by pseudomonic acid A
J. Bacteriol.
171
4518-4520
1989
Bacillus subtilis (P13956)
Zhong, P.; Pratt, S.D.; Edalji, R.P.; Walter, K.A.; Holzman, T.F.; Shivakumar, A.G.; Katz, L.
Substrate requirements for ErmC' methyltransferase activity
J. Bacteriol.
177
4327-4332
1995
Bacillus subtilis (P13956), Bacillus subtilis
Schluckebier, G.; Zhong, P.; Stewart, K.D.
Kavanaugh, T.J.; Abad-Zapatero, C.: The 2.2 A structure of the rRNA methyltransferase ErmC' and its complexes with cofactor and cofactor analogs: implications for the reaction mechanism
J. Mol. Biol.
289
277-291
1999
Bacillus subtilis (P13956)
Maravic, G.; Bujnicki, J.M.; Feder, M.; Pongor, S.; Flgel, M.
Alanine-scanning mutagenesis of the predicted rRNA-binding domain of ErmC' redefines the substrate-binding site and suggests a model for protein-RNA interactions
Nucleic Acids Res.
31
4941-4949
2003
Bacillus subtilis (P13956), Bacillus subtilis BD1167 (P13956)
EXTERNAL LINKS (specific for EC-Number 2.1.1.184)
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2023.1 (January 2023)
Legal
Curated at
Member of
