BRENDA - Enzyme Database
show all sequences of 2.1.1.224

High-level expression and reconstitution of active Cfr, a radical-SAM rRNA methyltransferase that confers resistance to ribosome-acting antibiotics

Booth, M.P.; Challand, M.R.; Emery, D.C.; Roach, P.L.; Spencer, J.; Protein Expr. Purif. 74, 204-210 (2010)

Data extracted from this reference:

Activating Compound
Activating Compound
Commentary
Organism
Structure
Sodium dithionite
-
Azotobacter vinelandii
Cloned(Commentary)
Cloned (Commentary)
Organism
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
Azotobacter vinelandii
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
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
Azotobacter vinelandii
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
Azotobacter vinelandii
the enzyme methylates the 8 position of 23S rRNA residue A2503 to confer resistance to multiple antibiotic classes acting upon the large subunit of the bacterial ribosome
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
-
-
?
Organism
Organism
UniProt
Commentary
Textmining
Azotobacter vinelandii
-
isc operon, gene cfr
-
Purification (Commentary)
Purification (Commentary)
Organism
recombinant His6-tagged Cfr from Escherichia coli strain BL21Star by nickel affinity chromatography and gel filtration
Azotobacter vinelandii
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
the enzyme methylates the 8 position of 23S rRNA residue A2503 to confer resistance to multiple antibiotic classes acting upon the large subunit of the bacterial ribosome
720968
Azotobacter vinelandii
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA
in the presence of sodium dithionite reconstituted Cfr is both reducible and able to cleave S-adenosyl-L-methionine to 5'-deoxyadeonsine, DOA
720968
Azotobacter vinelandii
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
-
-
-
?
Subunits
Subunits
Commentary
Organism
monomer
Cfr in solution, secondary structure analysis, circular dichroism spectroscopy and analytical gel filtration, overview
Azotobacter vinelandii
Synonyms
Synonyms
Commentary
Organism
Cfr
-
Azotobacter vinelandii
radical-S-adenosyl-L-methionine enzyme
-
Azotobacter vinelandii
radical-SAM enzyme
-
Azotobacter vinelandii
radical-SAM rRNA methyltransferase
-
Azotobacter vinelandii
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
37
-
assay at
Azotobacter vinelandii
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
9.5
-
assay at
Azotobacter vinelandii
Cofactor
Cofactor
Commentary
Organism
Structure
S-adenosyl-L-methionine
-
Azotobacter vinelandii
Activating Compound (protein specific)
Activating Compound
Commentary
Organism
Structure
Sodium dithionite
-
Azotobacter vinelandii
Cloned(Commentary) (protein specific)
Commentary
Organism
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
Azotobacter vinelandii
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
S-adenosyl-L-methionine
-
Azotobacter vinelandii
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
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
Azotobacter vinelandii
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
Azotobacter vinelandii
the enzyme methylates the 8 position of 23S rRNA residue A2503 to confer resistance to multiple antibiotic classes acting upon the large subunit of the bacterial ribosome
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
recombinant His6-tagged Cfr from Escherichia coli strain BL21Star by nickel affinity chromatography and gel filtration
Azotobacter vinelandii
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
the enzyme methylates the 8 position of 23S rRNA residue A2503 to confer resistance to multiple antibiotic classes acting upon the large subunit of the bacterial ribosome
720968
Azotobacter vinelandii
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
-
-
-
?
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA
in the presence of sodium dithionite reconstituted Cfr is both reducible and able to cleave S-adenosyl-L-methionine to 5'-deoxyadeonsine, DOA
720968
Azotobacter vinelandii
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
monomer
Cfr in solution, secondary structure analysis, circular dichroism spectroscopy and analytical gel filtration, overview
Azotobacter vinelandii
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
37
-
assay at
Azotobacter vinelandii
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
9.5
-
assay at
Azotobacter vinelandii
General Information
General Information
Commentary
Organism
physiological function
the enzyme methylates the 8 position of 23S rRNA residue A2503 to confer resistance to multiple antibiotic classes acting upon the large subunit of the bacterial ribosome. 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
Azotobacter vinelandii
General Information (protein specific)
General Information
Commentary
Organism
physiological function
the enzyme methylates the 8 position of 23S rRNA residue A2503 to confer resistance to multiple antibiotic classes acting upon the large subunit of the bacterial ribosome. 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
Azotobacter vinelandii
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
-
-
2
-
2
-
-
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-
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2
3
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2
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1
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1
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3
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1
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2
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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2
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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
-
-
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
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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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1
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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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1
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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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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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13
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706726
Giessing
Identification of 8-methyladen ...
Escherichia coli K-12
RNA
15
327-336
2009
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701724
Long
The Cfr rRNA methyltransferase ...
Escherichia coli
Antimicrob. Agents Chemother.
50
2500-2505
2006
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705766
Kehrenberg
A new mechanism for chloramphe ...
Escherichia coli
Mol. Microbiol.
57
1064-1073
2005
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