Literature summary extracted from

Booth, M.P.; Challand, M.R.; Emery, D.C.; Roach, P.L.; Spencer, J.
High-level expression and reconstitution of active Cfr, a radical-SAM rRNA methyltransferase that confers resistance to ribosome-acting antibiotics (2010), Protein Expr. Purif., 74, 204-210.

Activating Compound

EC Number	Activating Compound	Comment	Organism	Structure
2.1.1.224	Sodium dithionite	-	Azotobacter vinelandii

Cloned(Commentary)

EC Number	Cloned (Comment)	Organism
2.1.1.224	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

EC Number	Metals/Ions	Comment	Organism	Structure
2.1.1.224	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)

EC Number	Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
2.1.1.224	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

EC Number	Organism	UniProt	Comment	Textmining
2.1.1.224	Azotobacter vinelandii	-	isc operon, gene cfr	-

Purification (Commentary)

EC Number	Purification (Comment)	Organism
2.1.1.224	recombinant His6-tagged Cfr from Escherichia coli strain BL21Star by nickel affinity chromatography and gel filtration	Azotobacter vinelandii

Substrates and Products (Substrate)

EC Number	Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
2.1.1.224	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	Azotobacter vinelandii	S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA	-	?
2.1.1.224	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	Azotobacter vinelandii	S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA	-	?

Subunits

EC Number	Subunits	Comment	Organism
2.1.1.224	monomer	Cfr in solution, secondary structure analysis, circular dichroism spectroscopy and analytical gel filtration, overview	Azotobacter vinelandii

Synonyms

EC Number	Synonyms	Comment	Organism
2.1.1.224	Cfr	-	Azotobacter vinelandii
2.1.1.224	radical-S-adenosyl-L-methionine enzyme	-	Azotobacter vinelandii
2.1.1.224	radical-SAM enzyme	-	Azotobacter vinelandii
2.1.1.224	radical-SAM rRNA methyltransferase	-	Azotobacter vinelandii

Temperature Optimum [°C]

EC Number	Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
2.1.1.224	37	-	assay at	Azotobacter vinelandii

pH Optimum

EC Number	pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
2.1.1.224	9.5	-	assay at	Azotobacter vinelandii

Cofactor

EC Number	Cofactor	Comment	Organism	Structure
2.1.1.224	S-adenosyl-L-methionine	-	Azotobacter vinelandii

General Information

EC Number	General Information	Comment	Organism
2.1.1.224	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

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Release 2023.1 (January 2023)