BRENDA - Enzyme Database
show all sequences of 2.1.1.224

RlmN and Cfr are radical SAM enzymes involved in methylation of ribosomal RNA

Yan, F.; LaMarre, J.M.; Röhrich, R.; Wiesner, J.; Jomaa, H.; Mankin, A.S.; Fujimori, D.G.; J. Am. Chem. Soc. 132, 3953-3964 (2010)

Data extracted from this reference:

Cloned(Commentary)
Cloned (Commentary)
Organism
expressed as C-terminally His6-tagged enzyme, wild-type and mutant enzymes C125A, C129A and C132A
Escherichia coli
Engineering
Protein Variants
Commentary
Organism
C112A
inactive mutant enzyme
Escherichia coli
C116A
inactive mutant enzyme
Escherichia coli
C119A
inactive mutant enzyme
Escherichia coli
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Iron
the enzyme requires an intact [4Fe-S] cluster for catalysis
Escherichia coli
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
37000
-
x * 37000, SDS-PAGE
Escherichia coli
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA
Escherichia coli
Cfr is an plasmid-acquired methyltransferase that protects cells from the action of antibiotics
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
-
-
?
Organism
Organism
UniProt
Commentary
Textmining
Escherichia coli
-
plasmid(pMS2)-encoded enzyme
-
Purification (Commentary)
Purification (Commentary)
Organism
C-terminally His6-tagged wild-type enzyme and mutant enzymes C112A, C116A and C119A
Escherichia coli
Reaction
Reaction
Commentary
Organism
Reaction ID
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
the enzyme catalyzes adenosine methylation by using a radical mechanism for substrate activation. The radical chemistry is enabled by the [4Fe-4S] cluster
Escherichia coli
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
2 S-adenosyl-L-methionine + 2-methyladenine2503 in 23S rRNA
S-adenosylmethionine is both the methyl donor and the source of a 5'-deoxyadenosyl radical, which activates the substrate for methylation
704203
Escherichia coli
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 2,8-dimethyladenine2503 in 23S rRNA
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
Cfr is an plasmid-acquired methyltransferase that protects cells from the action of antibiotics
704203
Escherichia coli
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA
S-adenosylmethionine is both the methyl donor and the source of a 5'-deoxyadenosyl radical, which activates the substrate for methylation. The enzyme can utilize protein-free 23S rRNA as a substrate, but not the fully assembled large ribosomal subunit, suggesting that the methylations take place during the assembly of the ribosome. The key recognition elements in the 23S rRNA are helices 90-92 and the adjacent single stranded RNA that encompasses A2503
704203
Escherichia coli
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
identification of the reaction products as 8-methyladenine2503, and 2,8-dimethyladenine2503
-
-
?
Subunits
Subunits
Commentary
Organism
?
x * 37000, SDS-PAGE
Escherichia coli
Synonyms
Synonyms
Commentary
Organism
Cfr
-
Escherichia coli
Cloned(Commentary) (protein specific)
Commentary
Organism
expressed as C-terminally His6-tagged enzyme, wild-type and mutant enzymes C125A, C129A and C132A
Escherichia coli
Engineering (protein specific)
Protein Variants
Commentary
Organism
C112A
inactive mutant enzyme
Escherichia coli
C116A
inactive mutant enzyme
Escherichia coli
C119A
inactive mutant enzyme
Escherichia coli
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Iron
the enzyme requires an intact [4Fe-S] cluster for catalysis
Escherichia coli
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
37000
-
x * 37000, SDS-PAGE
Escherichia coli
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA
Escherichia coli
Cfr is an plasmid-acquired methyltransferase that protects cells from the action of antibiotics
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
C-terminally His6-tagged wild-type enzyme and mutant enzymes C112A, C116A and C119A
Escherichia coli
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
2 S-adenosyl-L-methionine + 2-methyladenine2503 in 23S rRNA
S-adenosylmethionine is both the methyl donor and the source of a 5'-deoxyadenosyl radical, which activates the substrate for methylation
704203
Escherichia coli
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 2,8-dimethyladenine2503 in 23S rRNA
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
Cfr is an plasmid-acquired methyltransferase that protects cells from the action of antibiotics
704203
Escherichia coli
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA
S-adenosylmethionine is both the methyl donor and the source of a 5'-deoxyadenosyl radical, which activates the substrate for methylation. The enzyme can utilize protein-free 23S rRNA as a substrate, but not the fully assembled large ribosomal subunit, suggesting that the methylations take place during the assembly of the ribosome. The key recognition elements in the 23S rRNA are helices 90-92 and the adjacent single stranded RNA that encompasses A2503
704203
Escherichia coli
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
identification of the reaction products as 8-methyladenine2503, and 2,8-dimethyladenine2503
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
?
x * 37000, SDS-PAGE
Escherichia coli
Other publictions for EC 2.1.1.224
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Synonyms
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
735450
Hansen
A cfr-like gene from Clostridi ...
Clostridioides difficile
Antimicrob. Agents Chemother.
59
5841-5843
2015
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1
1
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733111
Atkinson
Distinction between the Cfr me ...
Staphylococcus sciuri
Antimicrob. Agents Chemother.
57
4019-4026
2013
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1
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1
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1
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1
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2
2
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735095
Challand
Cysteine methylation controls ...
Staphylococcus aureus, Staphylococcus sciuri
PLoS ONE
8
e67979
2013
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2
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2
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3
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2
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1
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2
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736280
Wang
-
Quantum chemistry studies of a ...
Staphylococcus aureus
Int. J. Quantum Chem.
113
1409-1415
2013
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1
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3
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718598
Locke
Genetic environment and stabil ...
Staphylococcus aureus, Staphylococcus aureus CM05
Antimicrob. Agents Chemother.
56
332-340
2012
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1
-
1
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5
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718601
Hansen
The order Bacillales hosts fun ...
Bacillus amyloliquefaciens, Bacillus amyloliquefaciens FZB42, Bacillus clausii, Brevibacillus brevis, Brevibacillus brevis NBRC 100599
Antimicrob. Agents Chemother.
56
3563-3567
2012
-
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3
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5
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10
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10
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3
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3
3
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5
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10
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6
6
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716783
Yan
RNA methylation by radical SAM ...
Escherichia coli
Proc. Natl. Acad. Sci. USA
108
3930-3934
2011
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716926
Boal
Structural basis for methyl tr ...
Escherichia coli
Science
332
1089-1092
2011
-
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716927
Grove
A radically different mechanis ...
Escherichia coli
Science
332
604-607
2011
-
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1
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718587
LaMarre
Low fitness cost of the multid ...
Staphylococcus aureus, Staphylococcus aureus RN4220
Antimicrob. Agents Chemother.
55
3714-3719
2011
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1
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1
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1
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2
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21
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1
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2
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704203
Yan
RlmN and Cfr are radical SAM e ...
Escherichia coli
J. Am. Chem. Soc.
132
3953-3964
2010
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1
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3
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1
1
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720968
Booth
High-level expression and reco ...
Azotobacter vinelandii
Protein Expr. Purif.
74
204-210
2010
1
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1
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1
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705983
Kaminska
Insights into the structure, f ...
Escherichia coli
Nucleic Acids Res.
38
1652-1663
2009
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1
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13
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1
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13
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1
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706726
Giessing
Identification of 8-methyladen ...
Escherichia coli K-12
RNA
15
327-336
2009
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4
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701724
Long
The Cfr rRNA methyltransferase ...
Escherichia coli
Antimicrob. Agents Chemother.
50
2500-2505
2006
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1
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705766
Kehrenberg
A new mechanism for chloramphe ...
Escherichia coli
Mol. Microbiol.
57
1064-1073
2005
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