BRENDA - Enzyme Database

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)
EC Number
Commentary
Organism
2.1.1.192
expressed in Escherichia coli Rosetta2(DE3)pLysS cells
Staphylococcus sciuri
2.1.1.224
expressed in Escherichia coli Rosetta2(DE3)pLysS cells
Staphylococcus sciuri
2.1.1.224
gene cfr, recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain +Rosetta2(DE3)
Staphylococcus aureus
Engineering
EC Number
Amino acid exchange
Commentary
Organism
2.1.1.192
C338A
the mutant binds S-adenosyl-L-methionine with wild type affinity, while oxidation of the [4Fe-4S] cluster is not observed
Staphylococcus sciuri
2.1.1.224
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
2.1.1.224
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]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.1.1.192
41085
-
x * 41085, electrospray mass spectrometry
Staphylococcus sciuri
2.1.1.192
41088
-
x * 41088, fully methylated protein, calculated from amino acid sequence
Staphylococcus sciuri
2.1.1.224
41085
-
x * 41085, electrospray mass spectrometry
Staphylococcus sciuri
2.1.1.224
41088
-
x * 41088, fully methylated protein, calculated from amino acid sequence
Staphylococcus sciuri
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.1.1.192
S-adenosyl-L-methionine + adenine2503 in 23S rRNA
Staphylococcus sciuri
-
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 2-methyladenine2503 in 23S rRNA
-
-
?
2.1.1.224
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
-
-
?
2.1.1.224
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
?
-
-
-
2.1.1.224
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
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.1.1.192
Staphylococcus sciuri
Q9FBG4
-
-
2.1.1.224
Staphylococcus aureus
-
methicillin-resistant, gene cfr
-
2.1.1.224
Staphylococcus sciuri
Q9FBG4
-
-
Reaction
EC Number
Reaction
Commentary
Organism
2.1.1.224
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)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.1.1.192
S-adenosyl-L-methionine + adenine2503 in 23S rRNA
-
735095
Staphylococcus sciuri
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 2-methyladenine2503 in 23S rRNA
-
-
-
?
2.1.1.224
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
-
-
-
?
2.1.1.224
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
-
-
-
?
2.1.1.224
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
?
-
-
-
-
2.1.1.224
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
EC Number
Subunits
Commentary
Organism
2.1.1.192
?
x * 41085, electrospray mass spectrometry; x * 41088, fully methylated protein, calculated from amino acid sequence
Staphylococcus sciuri
2.1.1.224
?
x * 41085, electrospray mass spectrometry; x * 41088, fully methylated protein, calculated from amino acid sequence
Staphylococcus sciuri
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
2.1.1.192
[4Fe-4S]-center
the enzyme contains a [4Fe-4S] cluster
Staphylococcus sciuri
2.1.1.224
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
2.1.1.224
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
2.1.1.224
[4Fe-4S]-center
the enzyme contains a [4Fe-4S] cluster
Staphylococcus sciuri
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.1.1.192
expressed in Escherichia coli Rosetta2(DE3)pLysS cells
Staphylococcus sciuri
2.1.1.224
expressed in Escherichia coli Rosetta2(DE3)pLysS cells
Staphylococcus sciuri
2.1.1.224
gene cfr, recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain +Rosetta2(DE3)
Staphylococcus aureus
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
2.1.1.192
[4Fe-4S]-center
the enzyme contains a [4Fe-4S] cluster
Staphylococcus sciuri
2.1.1.224
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
2.1.1.224
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
2.1.1.224
[4Fe-4S]-center
the enzyme contains a [4Fe-4S] cluster
Staphylococcus sciuri
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
2.1.1.192
C338A
the mutant binds S-adenosyl-L-methionine with wild type affinity, while oxidation of the [4Fe-4S] cluster is not observed
Staphylococcus sciuri
2.1.1.224
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
2.1.1.224
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)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.1.1.192
41085
-
x * 41085, electrospray mass spectrometry
Staphylococcus sciuri
2.1.1.192
41088
-
x * 41088, fully methylated protein, calculated from amino acid sequence
Staphylococcus sciuri
2.1.1.224
41085
-
x * 41085, electrospray mass spectrometry
Staphylococcus sciuri
2.1.1.224
41088
-
x * 41088, fully methylated protein, calculated from amino acid sequence
Staphylococcus sciuri
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.1.1.192
S-adenosyl-L-methionine + adenine2503 in 23S rRNA
Staphylococcus sciuri
-
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 2-methyladenine2503 in 23S rRNA
-
-
?
2.1.1.224
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
-
-
?
2.1.1.224
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
?
-
-
-
2.1.1.224
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)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.1.1.192
S-adenosyl-L-methionine + adenine2503 in 23S rRNA
-
735095
Staphylococcus sciuri
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 2-methyladenine2503 in 23S rRNA
-
-
-
?
2.1.1.224
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
-
-
-
?
2.1.1.224
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
-
-
-
?
2.1.1.224
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
?
-
-
-
-
2.1.1.224
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)
EC Number
Subunits
Commentary
Organism
2.1.1.192
?
x * 41085, electrospray mass spectrometry; x * 41088, fully methylated protein, calculated from amino acid sequence
Staphylococcus sciuri
2.1.1.224
?
x * 41085, electrospray mass spectrometry; x * 41088, fully methylated protein, calculated from amino acid sequence
Staphylococcus sciuri
General Information
EC Number
General Information
Commentary
Organism
2.1.1.224
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
2.1.1.224
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)
EC Number
General Information
Commentary
Organism
2.1.1.224
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
2.1.1.224
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