Literature summary for 2.1.1.224 extracted from

Yan, F.; LaMarre, J.M.; R�hrich, R.; Wiesner, J.; Jomaa, H.; Mankin, A.S.; Fujimori, D.G.
RlmN and Cfr are radical SAM enzymes involved in methylation of ribosomal RNA (2010), J. Am. Chem. Soc., 132, 3953-3964.

Search for more literature for 2.1.1.224

Cloned(Commentary)

Cloned (Comment)	Organism
expressed as C-terminally His6-tagged enzyme, wild-type and mutant enzymes C125A, C129A and C132A	Escherichia coli

Protein Variants

Protein Variants	Comment	Organism
C112A	inactive mutant enzyme	Escherichia coli
C116A	inactive mutant enzyme	Escherichia coli
C119A	inactive mutant enzyme	Escherichia coli

Metals/Ions

Metals/Ions	Comment	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]	Comment	Organism
37000	-	x * 37000, SDS-PAGE	Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
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	Comment	Textmining
Escherichia coli	-	plasmid(pMS2)-encoded enzyme	-

Purification (Commentary)

Purification (Comment)	Organism
C-terminally His6-tagged wild-type enzyme and mutant enzymes C112A, C116A and C119A	Escherichia coli

Reaction

Reaction	Comment	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	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
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	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	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	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	Comment	Organism
?	x * 37000, SDS-PAGE	Escherichia coli

Synonyms

Synonyms	Comment	Organism
Cfr	-	Escherichia coli

Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.

Release 2023.1 (January 2023)