BRENDA - Enzyme Database
show all sequences of 2.1.1.224

Cysteine methylation controls radical generation in the Cfr radical AdoMet rRNA methyltransferase

Challand, M.R.; Salvadori, E.; Driesener, R.C.; Kay, C.W.; Roach, P.L.; Spencer, J.; PLoS ONE 8, e67979 (2013)

Data extracted from this reference:

Cloned(Commentary)
Cloned (Commentary)
Organism
expressed in Escherichia coli Rosetta2(DE3)pLysS cells
Staphylococcus sciuri
gene cfr, recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain +Rosetta2(DE3)
Staphylococcus aureus
Engineering
Protein Variants
Commentary
Organism
C338A
site-directed mutagenesis, Cys338Ala Cfr binds S-adenosyl-L-methionine with equivalent affinity, oxidation of the [4Fe-4S] cluster is not observed
Staphylococcus aureus
C338A
the mutant binds S-adenosyl-L-methionine with wild type affinity, while oxidation of the [4Fe-4S] cluster is not observed
Staphylococcus sciuri
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
41085
-
x * 41085, electrospray mass spectrometry
Staphylococcus sciuri
41088
-
x * 41088, fully methylated protein, calculated from amino acid sequence
Staphylococcus sciuri
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 + 2 reduced [4Fe-4S] ferredoxin
Staphylococcus aureus
-
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [4Fe-4S] ferredoxin
-
-
?
additional information
Staphylococcus aureus
enzyme Cfr methylates adenosine 2503 of the 23S rRNA in the peptidyltransferase centre (P-site) of the bacterial ribosome. In wild-type Cfr, where Cys338 is methylated, S-adenosyl-L-methionine binding leads to rapid oxidation of the [4Fe-4S] cluster and production of 5'-deoxyadenosine
?
-
-
-
S-adenosyl-L-methionine + adenine2503 in 23S rRNA + reduced [2Fe-2S] ferredoxin
Staphylococcus sciuri
-
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + oxidized [2Fe-2S] ferredoxin
-
-
-
Organism
Organism
UniProt
Commentary
Textmining
Staphylococcus aureus
-
methicillin-resistant, gene cfr
-
Staphylococcus sciuri
Q9FBG4
-
-
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
enzyme Cfr consumes two AdoMet equivalents per reaction cycle to support both methyl transfer to Cfr Cys338 (AdoMet1, step 1) and subsequently generation of the 59dA. radical (AdoMet2, step2)
Staphylococcus aureus
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 + adenine2503 in 23S rRNA + 2 reduced [4Fe-4S] ferredoxin
-
735095
Staphylococcus aureus
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [4Fe-4S] ferredoxin
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [4Fe-4S] ferredoxin
enzyme Cfr consumes two AdoMet equivalents per reaction cycle to support both methyl transfer to Cfr Cys338 (AdoMet1, step 1) and subsequently generation of the 59dA. radical (AdoMet2, step2)
735095
Staphylococcus aureus
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [4Fe-4S] ferredoxin
-
-
-
?
additional information
enzyme Cfr methylates adenosine 2503 of the 23S rRNA in the peptidyltransferase centre (P-site) of the bacterial ribosome. In wild-type Cfr, where Cys338 is methylated, S-adenosyl-L-methionine binding leads to rapid oxidation of the [4Fe-4S] cluster and production of 5'-deoxyadenosine
735095
Staphylococcus aureus
?
-
-
-
-
S-adenosyl-L-methionine + adenine2503 in 23S rRNA + reduced [2Fe-2S] ferredoxin
-
735095
Staphylococcus sciuri
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + oxidized [2Fe-2S] ferredoxin
-
-
-
-
Subunits
Subunits
Commentary
Organism
?
x * 41085, electrospray mass spectrometry; x * 41088, fully methylated protein, calculated from amino acid sequence
Staphylococcus sciuri
Synonyms
Synonyms
Commentary
Organism
Cfr
-
Staphylococcus aureus
Cfr
-
Staphylococcus sciuri
radical AdoMet rRNA methyltransferase
-
Staphylococcus sciuri
Cofactor
Cofactor
Commentary
Organism
Structure
Ferredoxin
enzyme Cfr contains a single [4Fe-4S] cluster that binds two separate molecules of S-adenosyl-L-methionine during the reaction cycle
Staphylococcus aureus
S-adenosyl-L-methionine
enzyme Cfr contains a single [4Fe-4S] cluster that binds two separate molecules of S-adenosyl-L-methionine during the reaction cycle
Staphylococcus aureus
[4Fe-4S]-center
the enzyme contains a [4Fe-4S] cluster
Staphylococcus sciuri
Cloned(Commentary) (protein specific)
Commentary
Organism
expressed in Escherichia coli Rosetta2(DE3)pLysS cells
Staphylococcus sciuri
gene cfr, recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain +Rosetta2(DE3)
Staphylococcus aureus
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
Ferredoxin
enzyme Cfr contains a single [4Fe-4S] cluster that binds two separate molecules of S-adenosyl-L-methionine during the reaction cycle
Staphylococcus aureus
S-adenosyl-L-methionine
enzyme Cfr contains a single [4Fe-4S] cluster that binds two separate molecules of S-adenosyl-L-methionine during the reaction cycle
Staphylococcus aureus
[4Fe-4S]-center
the enzyme contains a [4Fe-4S] cluster
Staphylococcus sciuri
Engineering (protein specific)
Protein Variants
Commentary
Organism
C338A
site-directed mutagenesis, Cys338Ala Cfr binds S-adenosyl-L-methionine with equivalent affinity, oxidation of the [4Fe-4S] cluster is not observed
Staphylococcus aureus
C338A
the mutant binds S-adenosyl-L-methionine with wild type affinity, while oxidation of the [4Fe-4S] cluster is not observed
Staphylococcus sciuri
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
41085
-
x * 41085, electrospray mass spectrometry
Staphylococcus sciuri
41088
-
x * 41088, fully methylated protein, calculated from amino acid sequence
Staphylococcus sciuri
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 + 2 reduced [4Fe-4S] ferredoxin
Staphylococcus aureus
-
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [4Fe-4S] ferredoxin
-
-
?
additional information
Staphylococcus aureus
enzyme Cfr methylates adenosine 2503 of the 23S rRNA in the peptidyltransferase centre (P-site) of the bacterial ribosome. In wild-type Cfr, where Cys338 is methylated, S-adenosyl-L-methionine binding leads to rapid oxidation of the [4Fe-4S] cluster and production of 5'-deoxyadenosine
?
-
-
-
S-adenosyl-L-methionine + adenine2503 in 23S rRNA + reduced [2Fe-2S] ferredoxin
Staphylococcus sciuri
-
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + oxidized [2Fe-2S] ferredoxin
-
-
-
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 + adenine2503 in 23S rRNA + 2 reduced [4Fe-4S] ferredoxin
-
735095
Staphylococcus aureus
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [4Fe-4S] ferredoxin
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [4Fe-4S] ferredoxin
enzyme Cfr consumes two AdoMet equivalents per reaction cycle to support both methyl transfer to Cfr Cys338 (AdoMet1, step 1) and subsequently generation of the 59dA. radical (AdoMet2, step2)
735095
Staphylococcus aureus
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [4Fe-4S] ferredoxin
-
-
-
?
additional information
enzyme Cfr methylates adenosine 2503 of the 23S rRNA in the peptidyltransferase centre (P-site) of the bacterial ribosome. In wild-type Cfr, where Cys338 is methylated, S-adenosyl-L-methionine binding leads to rapid oxidation of the [4Fe-4S] cluster and production of 5'-deoxyadenosine
735095
Staphylococcus aureus
?
-
-
-
-
S-adenosyl-L-methionine + adenine2503 in 23S rRNA + reduced [2Fe-2S] ferredoxin
-
735095
Staphylococcus sciuri
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + oxidized [2Fe-2S] ferredoxin
-
-
-
-
Subunits (protein specific)
Subunits
Commentary
Organism
?
x * 41085, electrospray mass spectrometry; x * 41088, fully methylated protein, calculated from amino acid sequence
Staphylococcus sciuri
General Information
General Information
Commentary
Organism
additional information
the presence of a methyl group on Cfr Cys338 is a key determinant of the activity of the enzyme towards S-adenosyl-L-methionine, thus enabling a single active site to support two distinct modes of S-adenosyl-L-methionine cleavage
Staphylococcus aureus
physiological function
enzyme Cfr methylates adenosine 2503 of the 23S rRNA in the peptidyltransferase centre (P-site) of the bacterial ribosome. This modification protects host bacteria, notably methicillin-resistant Staphylococcus aureus (MRSA), from numerous antibiotics, including agents (e.g. linezolid, retapamulin)
Staphylococcus aureus
General Information (protein specific)
General Information
Commentary
Organism
additional information
the presence of a methyl group on Cfr Cys338 is a key determinant of the activity of the enzyme towards S-adenosyl-L-methionine, thus enabling a single active site to support two distinct modes of S-adenosyl-L-methionine cleavage
Staphylococcus aureus
physiological function
enzyme Cfr methylates adenosine 2503 of the 23S rRNA in the peptidyltransferase centre (P-site) of the bacterial ribosome. This modification protects host bacteria, notably methicillin-resistant Staphylococcus aureus (MRSA), from numerous antibiotics, including agents (e.g. linezolid, retapamulin)
Staphylococcus aureus
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
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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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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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2
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2
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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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1
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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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-
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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
-
10
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10
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3
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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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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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21
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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
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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
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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1
1
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701724
Long
The Cfr rRNA methyltransferase ...
Escherichia coli
Antimicrob. Agents Chemother.
50
2500-2505
2006
-
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7
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2
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705766
Kehrenberg
A new mechanism for chloramphe ...
Escherichia coli
Mol. Microbiol.
57
1064-1073
2005
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