Cloned (Comment) | Organism |
---|---|
recombinant expression of N-terminally His6-tagged wild-type and mutant enzymes, with additional FLAG and HA tags, in Escherichia coli | Streptomyces actuosus |
Protein Variants | Comment | Organism |
---|---|---|
R135A | site-directed mutagenesis, the inactive R135A mutant homodimer binds to the RNA causing a shift in electrophoretic mobility, and the heterodimer composed of active and inactive subunits also binds the RNA efficiently. The inactive heterodimer complex binds the RNA more efficiently than the inactive mutant homodimer | Streptomyces actuosus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Streptomyces actuosus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
S-adenosyl-L-methionine + adenosine1067 in 23S rRNA | Streptomyces actuosus | - |
S-adenosyl-L-homocysteine + 2'-O-methyladenosine1067 in 23S rRNA | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Streptomyces actuosus | P52391 | nosiheptide producer | - |
Purification (Comment) | Organism |
---|---|
recombinant N-terminally His6-tagged wild-type and mutant enzymes from Escherichia coli by nickel affinity chromatography and gel filtration | Streptomyces actuosus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | compared the methylation activity of the Arg135Ala NHR heterodimer with the wild-type NHR homodimer and a series of 29 nt wild-type and mutant RNA substrates. The inactive heterodimer complex binds the RNA more efficiently than the inactive mutant homodimer. Construction of diverse mutant RNA substrates in which A1067 is replaced by adenine structural analogues, A1067G retains significant activity, while 7-deaza-A and 1-methyl-A substitutions at A1067 reduce the activity, overview | Streptomyces actuosus | ? | - |
? | |
S-adenosyl-L-methionine + adenosine1067 in 23S rRNA | - |
Streptomyces actuosus | S-adenosyl-L-homocysteine + 2'-O-methyladenosine1067 in 23S rRNA | - |
? | |
S-adenosyl-L-methionine + adenosine1067 in 23S rRNA | as 58 nucleotide substrate for methylation, or as shortened 29 nt hairpin substrate, local base-base interactions play an important role in aligning the substrate 2'-hydroxyl group of A1067 for methyl group transfer, stoichiometry of RNA binding by the NHR dimer, overview | Streptomyces actuosus | S-adenosyl-L-homocysteine + 2'-O-methyladenosine1067 in 23S rRNA | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | SDS-PAGE and gel filtration | Streptomyces actuosus |
Synonyms | Comment | Organism |
---|---|---|
NHR | - |
Streptomyces actuosus |
NHR protein | - |
Streptomyces actuosus |
nosiheptide resistance methyltransferase | - |
Streptomyces actuosus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Streptomyces actuosus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Streptomyces actuosus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
S-adenosyl-L-methionine | - |
Streptomyces actuosus |
General Information | Comment | Organism |
---|---|---|
additional information | the NHR heterodimer requires only one functional subunit for RNA recognition and enzymatic activity. The catalytic residue is Arg135 | Streptomyces actuosus |
physiological function | the nosiheptide producer Streptomyces actuosis prevents self-intoxication by expressing the enzyme, which methylates the 2'-hydroxyl of 23 S rRNA nucleotide adenosine 1067 within the antibiotic binding site. Methylation at A1067 blocks thiazole binding directly. This region of 23S rRNA is also the binding site for the ribosomal protein L11 | Streptomyces actuosus |