Information on EC 2.1.1.224 - 23S rRNA (adenine2503-C8)-methyltransferase

Word Map on EC 2.1.1.224
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)

The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
2.1.1.224
-
RECOMMENDED NAME
GeneOntology No.
23S rRNA (adenine2503-C8)-methyltransferase
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin = S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin
show the reaction diagram
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
linezolid resistance
-
-
SYSTEMATIC NAME
IUBMB Comments
S-adenosyl-L-methionine:23S rRNA (adenine2503-C8)-methyltransferase
This enzyme is a member of the 'AdoMet radical' (radical SAM) family. S-Adenosyl-L-methionine acts as both a radical generator and as the source of the appended methyl group. It contains an [4Fe-S] cluster [1]. Cfr is an plasmid-acquired methyltransferase that protects cells from the action of antibiotics [1]. The enzyme methylates adenosine at position 2503 of 23S rRNA by a radical mechanism, transferring a CH2 group from S-adenosyl-L-methionine while retaining the hydrogen at the C-8 position of the adenine. Cfr first transfers an CH2 group to a conserved cysteine (Cys338 in Staphylococcus aureus) [7], the generated radical from a second S-adenosyl-L-methionine then attacks the methyl group, exctracting a hydrogen. The formed radical forms a covalent intermediate with the adenine group of the tRNA [8]. The enzyme will also methylate 2-methyladenine produced by the action of EC 2.1.1.192 [23S rRNA (adenine2503-C2)-methyltransferase].
CAS REGISTRY NUMBER
COMMENTARY hide
39369-30-7
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
isc operon, gene cfr
-
-
Manually annotated by BRENDA team
gene clbA, a cfr-like gene
-
-
Manually annotated by BRENDA team
gene clbA, a cfr-like gene
-
-
Manually annotated by BRENDA team
or strain domuvar, gene clbC, a cfr-like gene
UniProt
Manually annotated by BRENDA team
or strain domuvar, gene clbC, a cfr-like gene
UniProt
Manually annotated by BRENDA team
gene clbB, a cfr-like gene
UniProt
Manually annotated by BRENDA team
gene clbB, a cfr-like gene
UniProt
Manually annotated by BRENDA team
Peptoclostridium difficile
formerly Clostridium difficile, gene cfr
UniProt
Manually annotated by BRENDA team
gene cfr, while cfr is typically plasmid borne, in CM05 it is located on the chromosome and is coexpressed with ermB as part of the mlr operon
UniProt
Manually annotated by BRENDA team
gene cfr
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2 S-adenosyl-L-methionine + 2-methyladenine2503 in 23S rRNA
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 2,8-dimethyladenine2503 in 23S rRNA
show the reaction diagram
-
S-adenosylmethionine is both the methyl donor and the source of a 5'-deoxyadenosyl radical, which activates the substrate for methylation
incubation of Cfr with the wild-type 23S rRNA, already modified at the C2 position by the endogenous RlmN, provides 2,8-dimethyladenosine as the sole product
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
show the reaction diagram
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [4Fe-4S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [4Fe-4S] ferredoxin
show the reaction diagram
S-adenosyl-L-methionine + adenine2503 in 23S rRNA + reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + oxidized [2Fe-2S] ferredoxin
show the reaction diagram
-
-
-
-
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
show the reaction diagram
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [4Fe-4S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [4Fe-4S] ferredoxin
show the reaction diagram
S-adenosyl-L-methionine + adenine2503 in 23S rRNA + reduced [2Fe-2S] ferredoxin
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + oxidized [2Fe-2S] ferredoxin
show the reaction diagram
Q9FBG4
-
-
-
-
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Ferredoxin
-
S-adenosyl-L-methionine
[4Fe-4S]-center
the enzyme contains a [4Fe-4S] cluster
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Fe-S-clusters
-
contains a 4Fe-4S cluster. Radical-SAM enzymes contain a [4Fe-4S]+ cluster that is coordinated by the three conserved cysteine thiolate side chains in the CX3CX2C motif and one molecule of S-adenosyl-L-methionine
Fe2+
-
4Fe-4S cluster. Fe-S clusters are of structural, as well as functional, importance to Cfr. Radical-SAM enzymes use an Fe-S cluster to generate the 5'-deoxyadenosyl radical from SAM, enabling them to modify intrinsically unreactive centres such as adenosine C8. Anaerobic purification from Azotobacter vinelandii Cfr
Iron
-
the enzyme requires an intact [4Fe-S] cluster for catalysis
Mg2+
-
required
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Sodium dithionite
-
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
additional information
energy profile and thermodynamics, overview
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.5
-
assay at
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
additional information
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
-
Cfr in solution, secondary structure analysis, circular dichroism spectroscopy and analytical gel filtration, overview
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
C-terminally His6-tagged wild-type enzyme and mutant enzymes C112A, C116A and C119A
-
recombinant His6-tagged Cfr from Escherichia coli strain BL21Star by nickel affinity chromatography and gel filtration
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed as C-terminally His6-tagged enzyme, wild-type and mutant enzymes C125A, C129A and C132A
-
expressed in Escherichia coli Rosetta2(DE3)pLysS cells
gene cfr, operon isc, co-expression with isc proteins, expression of His6-tagged Cfr under control of an arabinose-inducible promoter in Escherichia coli strain BL21Star
-
gene cfr, recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain +Rosetta2(DE3)
-
gene cfr, sequence comparisons and phylogenetic analysis, primer extension analysis, cloning and expression in Escherichia coli strains TOP10 and AS19
gene cfr, The Cfr-like protein in Peptoclostridium difficile is hosted by a transposon, and the same gene is found in other strains, sequence comparisons, overview. Recombinant expression in Escherichia coli strain AS19 which then shows elevated MIC values for five classes of antibiotics, and recombinant expression in rlmN-deficient Escherichia coli strain JW2505-1
Peptoclostridium difficile
gene cfr, while cfr is typically plasmid borne, in CM05 it is located on the chromosome and is coexpressed with ermB as part of the mlr operon. DNA and amino acid sequence determination and analysis and genetic organization, overview. The cfr-containing mlr operon is located within a 15.5-kb plasmid-like insertion into 23S rRNA allele 4. In the LZDs colonies, designated CM05DELTA, a recombination event involving the two ermB genes had occurred, resulting in the deletion of cfr and the 3= flanking region, cfr-istAS-istBS-ermB
gene clbA, DNA and amino acid sequence determination and analysis, phylogenetic analysis, expression in Escherichia coli strain AS19 using plasmid pLJ10
-
gene clbB, DNA and amino acid sequence determination and analysis, phylogenetic analysis, expression in Escherichia coli strain AS19 using plasmid pLJ10
gene clbC, DNA and amino acid sequence determination and analysis, phylogenetic analysis, expression in Escherichia coli strain AS19 using plasmid pLJ10
gene gfr, ermB and cfr genes are naturally coexpressed under the Perm promoter in the mlr operon, expression of mlr results in modification of A2058 and A2503 in 23S rRNA and renders cells resistant to all clinically relevant antibiotics that target the large ribosomal subunit
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C105A
-
mutation eliminates the resistance to both florfenicol and tiamulin
C110A
-
mutation does not affect Cfr activity
C113A
no significant reduction in activity
C115A
mutation of the cysteines in the presumed radical S-adenosyl-L-methionine motif CxxxCxxC abolishes Cfr activity
C122A
mutation of the cysteines in the presumed radical S-adenosyl-L-methionine motif CxxxCxxC abolishes Cfr activity
C338A
-
mutation eliminates the resistance to both florfenicol and tiamulin
E91A
-
mutation eliminates the resistance to both florfenicol and tiamulin
F118A
-
mutation eliminates the resistance to both florfenicol and tiamulin
H214A
-
moderately decreased resistance to both florfenicol and tiamulin
Q28A
-
mutation eliminates the resistance to both florfenicol and tiamulin
R25A
-
mutation lowers the resistance to both florfenicol and tiamulin considerably
S189A
-
mutation lowers the resistance to both florfenicol and tiamulin considerably
S212A
-
mutation eliminates the resistance to both florfenicol and tiamulin
C119A
-
site-directed mutagenesis, the mutant Cfr is inactive, and mutant cells show no resistance against antibiotics
C338A
-
site-directed mutagenesis, Cys338Ala Cfr binds S-adenosyl-L-methionine with equivalent affinity, oxidation of the [4Fe-4S] cluster is not observed
C119A
-
site-directed mutagenesis, the mutant Cfr is inactive, and mutant cells show no resistance against antibiotics
-
C338A
the mutant binds S-adenosyl-L-methionine with wild type affinity, while oxidation of the [4Fe-4S] cluster is not observed
additional information
Show AA Sequence (106 entries)
Please use the Sequence Search for a certain query.