Cloned (Comment) | Organism |
---|---|
gene cfr, recombinant expression of wild-type and mutant enzymes in Escherichia coli strain JW2501-1 (rlmN knockout) and AS19 | Mammaliicoccus sciuri |
Crystallization (Comment) | Organism |
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homology modeling of structure and comparison with RlmN from Echerichia coli, EC 2.1.1.192 | Mammaliicoccus sciuri |
Protein Variants | Comment | Organism |
---|---|---|
additional information | construction of chimera between Cfr and RlmN from Echerichia coli to analyze C2/C8 and C2 methylation specificity, respectively. The catalytic site is expected to be responsible for the C2/C8 specificity. Almost all replacements show no function in the primer extension assay, apart from a few that have a weak effect | Mammaliicoccus sciuri |
additional information | construction of chimeric mutants of gene cfr, cfr1234567rrlmN and rlmN1234567rcfr, and construction of mixed genes containing gene fragment of cfr and rlmN genes, overview. Analysis of effects of the mutations on catalytic activity and antibiotics resistance | Mammaliicoccus sciuri |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin | Mammaliicoccus sciuri | - |
S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mammaliicoccus sciuri | Q9FBG4 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA | - |
Mammaliicoccus sciuri | 2 S-adenosyl-L-homocysteine + 2,8-dimethyladenine2503 in 23S rRNA | - |
? | |
2 S-adenosyl-L-methionine + adenine2503 in 23S rRNA + 2 reduced [2Fe-2S] ferredoxin | - |
Mammaliicoccus sciuri | S-adenosyl-L-homocysteine + L-methionine + 5'-deoxyadenosine + 8-methyladenine2503 in 23S rRNA + 2 oxidized [2Fe-2S] ferredoxin | - |
? | |
additional information | Cfr methylates A2503 of 23S rRNA at C8 and C2, while RlmN only performs a C2 methylation using the same mechanism of function | Mammaliicoccus sciuri | ? | - |
- |
Synonyms | Comment | Organism |
---|---|---|
Cfr | - |
Mammaliicoccus sciuri |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
4Fe-4S-center | - |
Mammaliicoccus sciuri | |
reduced [2Fe-2S] ferredoxin | - |
Mammaliicoccus sciuri | |
S-adenosyl-L-methionine | - |
Mammaliicoccus sciuri | |
[4Fe-4S] cluster | - |
Mammaliicoccus sciuri |
General Information | Comment | Organism |
---|---|---|
evolution | evolutionary relationship between the Cfr and RlmN (EC 2.1.1.192) enzymes, phylogenetic analysis, overview | Mammaliicoccus sciuri |
additional information | mechanisms of catalytic action of Cfr and related RlmN (EC 2.1.1.192), the methylation mechanism involves a transitory methylation of Cys338 for Cfr and Cys355 for RlmN, investigation of target binding to the active sites of the two enzymes, overview. Cfr and RlmN are methylated before transfer of the methyl group to the substrate. Homology structure modelling, molecular dynamics simulations, and calculation of the binding free energy, using structure of Escherichia coli RlmN (PDB ID 3RFA), the homology model is made with the [4Fe-4S] cluster and a SAM molecule positioned in the same way as seen in the RlmN X-ray structure. Defining regions of the active site to be interchanged to investigate C8/C2 specificity | Mammaliicoccus sciuri |
physiological function | antibiotic resistance effects of wild-type and mutant enzymes, overview | Mammaliicoccus sciuri |