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Literature summary for 3.1.26.4 extracted from

  • Jongruja, N.; You, D.J.; Angkawidjaja, C.; Kanaya, E.; Koga, Y.; Kanaya, S.
    Structure and characterization of RNase H3 from Aquifex aeolicus (2012), FEBS J., 279, 2737-2753.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli MIC2067(DE3) cells Aquifex aeolicus

Crystallization (Commentary)

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

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 [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

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/ Products (Substrates)

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

Organism UniProt Comment Textmining
Aquifex aeolicus O67644
-
-

Purification (Commentary)

Purification (Comment) Organism
HiTrap SP column chromatography and Hi-Trap heparin column chromatography Aquifex aeolicus

Substrates and Products (Substrate)

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

Synonyms Comment Organism
ribonuclease H3
-
Aquifex aeolicus
RNase H3
-
Aquifex aeolicus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
9
-
-
Aquifex aeolicus

pH Range

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