Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli MIC2067(DE3) cells | Aquifex aeolicus |
Crystallization (Comment) | Organism |
---|---|
sitting drop vapor diffusion method, using 0.2 m sodium citrate tribasic dehydrate, pH 8.3, and 20% (w/v) polyethylene glycol 3350, at 4°C | Aquifex aeolicus |
Protein Variants | Comment | Organism |
---|---|---|
E194A | the mutant exhibits 20 and 35% of the Mg2+- and Mn2+-dependent activities of the wild type enzyme, respectively | Aquifex aeolicus |
E196A | the mutant shows 84.8% Mn2+-dependent and 63% Mg2+-dependent activities compared to the wild type enzyme | Aquifex aeolicus |
E198A | the mutant shows 76.1% Mn2+-dependent and 63% Mg2+-dependent activities compared to the wild type enzyme | Aquifex aeolicus |
Y45A | the mutant shows 47.8% Mn2+-dependent and 5.4% Mg2+-dependent activities compared to the wild type enzyme | Aquifex aeolicus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0011 | - |
RNA-DNA hybrid | wild type enzyme, in 10 mM Tris/HCl (pH 9.0) containing 10 mM MgCl2, 100 mM KCl, 1 mM beta-mercaptoethanol and 0.05 mg/ml bovine serum albumin, at 30°C | Aquifex aeolicus | |
0.0012 | - |
RNA-DNA hybrid | wild type enzyme, in 10 mM Tris/HCl (pH 9.0) containing 10 mM MnCl2, 100 mM KCl, 1 mM beta-mercaptoethanol and 0.05 mg/ml bovine serum albumin, at 30°C | Aquifex aeolicus | |
0.0026 | - |
RNA-DNA hybrid | mutant enzyme Y45A, in 10 mM Tris/HCl (pH 9.0) containing 2 mM MnCl2, 100 mM KCl, 1 mM beta-mercaptoethanol and 0.05 mg/ml bovine serum albumin, at 30°C | Aquifex aeolicus | |
0.0028 | - |
RNA-DNA hybrid | mutant enzyme Y45A, in 10 mM Tris/HCl (pH 9.0) containing 10 mM MgCl2, 100 mM KCl, 1 mM beta-mercaptoethanol and 0.05 mg/ml bovine serum albumin, at 30°C | Aquifex aeolicus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
K+ | the enzyme exhibits the highest activity in the presence of 100 mm KCl | Aquifex aeolicus | |
Mg2+ | highest activity in the presence of 10 mM MgCl2 | Aquifex aeolicus | |
Mn2+ | highest activity in the presence of 5 mM MnCl2 | Aquifex aeolicus | |
additional information | the enzyme exhibits little activity (less than 0.002% of the maximal activity) in the presence of ZnCl2, CaCl2, CoCl2 and NiCl2 | Aquifex aeolicus | |
Na+ | the enzyme exhibits the highest activity in the presence of 100 mm NaCl | Aquifex aeolicus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
RNA-DNA hybrid + H2O | Aquifex aeolicus | the enzyme specifically cleaves the RNA strand of RNA/DNA hybrids | ribonucleotide 5'-phosphomonoester + ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Aquifex aeolicus | O67644 | - |
- |
Purification (Comment) | Organism |
---|---|
HiTrap SP column chromatography and Hi-Trap heparin column chromatography | Aquifex aeolicus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
RNA-DNA hybrid + H2O | the enzyme specifically cleaves the RNA strand of RNA/DNA hybrids | Aquifex aeolicus | ribonucleotide 5'-phosphomonoester + ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
ribonuclease H3 | - |
Aquifex aeolicus |
RNase H3 | - |
Aquifex aeolicus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
9 | - |
- |
Aquifex aeolicus |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
7 | 11 | the enzyme exhibits approximately 20-30% of maximal activity at pH 7.0 and 11.0 | Aquifex aeolicus |