Cloned (Comment) | Organism |
---|---|
gene rnc, overexpression of wild-type and mutants in strain BL21(DE3) as His-tagged proteins | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
E117D | site-directed mutagenesis, mutant exhibits normal homodimeric behaviour, can bind substrates but shows highly reduced hydrolysis activity compared to the wild-type enzyme | Escherichia coli |
E117Q | site-directed mutagenesis, mutant exhibits normal homodimeric behaviour, can bind substrates but is unable to cleave the substrates | Escherichia coli |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Mn2+ | at concentrations above 5 mM, mutant E117D is not inhibited, inhibition of the wild-type enzyme by excess Mn2+ cannot be relieved by high concentrations of Mg2+, inhibition mechanism | Escherichia coli | |
additional information | no inhibition by high concentrations of Mg2+ | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | catalytic and substrate binding kinetics | Escherichia coli | |
0.00011 | - |
R1.1 RNA | wild-type enzyme, pH 7.5, 37°C, in presence of Mn2+ | Escherichia coli | |
0.00034 | - |
R1.1 RNA | wild-type enzyme, pH 7.5, 37°C, in presence of Mg2+ | Escherichia coli | |
0.00064 | - |
R1.1 RNA | mutant E117D, pH 7.5, 37°C, in presence of Mn2+ | Escherichia coli | |
38 | - |
Mn2+ | mutant E117D, pH 7.5, 37°C | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Co2+ | can partially substitute for Mg2+ or Mn2+ | Escherichia coli | |
Mg2+ | required | Escherichia coli | |
Mn2+ | can substitute for Mg2+, showing slightly higher activity with the wild-type enzyme, highly activates the mutant E117D, no activity with mutant E117Q | Escherichia coli | |
additional information | no activity of mutant E117Q with Ca2+, Ni2+, Co2+, or Sr2+ | Escherichia coli | |
Ni2+ | can partially substitute for Mg2+ or Mn2+ | Escherichia coli |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
29000 | - |
2 * 29000, about, recombinant His-tagged enzymes, SDS-PAGE | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
dsRNA + H2O | Escherichia coli | enzyme is involved in the maturation and decay of cellular, phage and plasmid RNAs | mature RNA | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant HIs-tagged wild-type and mutant enzymes | Escherichia coli |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
Endonucleolytic cleavage to a 5'-phosphomonoester | reaction mechanism, conserved Mn2+-dependent Glu117 is required, overview | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
dsRNA + H2O | - |
Escherichia coli | mature RNA | - |
? | |
dsRNA + H2O | enzyme is involved in the maturation and decay of cellular, phage and plasmid RNAs | Escherichia coli | mature RNA | - |
? | |
R1.1 RNA + H2O | substrate is enzymatically synthesized based on the R1.1 processing signal, which is encoded in the phage T7 genetic early regionbetween genes 1.0 and 1.1, 1 cleavage site | Escherichia coli | 2 fragments of R1.1 RNA | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | 2 * 29000, about, recombinant His-tagged enzymes, SDS-PAGE | Escherichia coli |
More | the enzyme consists of a catalytic and a substrate binding domain | Escherichia coli |
Synonyms | Comment | Organism |
---|---|---|
RNase III | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00035 | - |
R1.1 RNA | mutant E117D, pH 7.5, 37°C, in presence of Mn2+ | Escherichia coli | |
0.023 | - |
R1.1 RNA | wild-type enzyme, pH 7.5, 37°C, in presence of Mn2+ | Escherichia coli | |
0.063 | - |
R1.1 RNA | wild-type enzyme, pH 7.5, 37°C, in presence of Mg2+ | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Escherichia coli |