Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutants enzymes in Escherichia coli BL21(DE3) | Methanocaldococcus jannaschii |
Protein Variants | Comment | Organism |
---|---|---|
D112N | site-directed mutagenesis, 0.17% activity compared to the wild-type enzyme, no Mn2+ binding | Methanocaldococcus jannaschii |
D149N | site-directed mutagenesis, 0.12% activity and slightly weakened Mn2+ binding properties compared to the wild-type enzyme | Methanocaldococcus jannaschii |
D7N | site-directed mutagenesis, 0.13% activity and slightly weakened Mn2+ binding properties compared to the wild-type enzyme | Methanocaldococcus jannaschii |
E8Q | site-directed mutagenesis, 0.14% activity and slightly weakened Mn2+ binding properties compared to the wild-type enzyme | Methanocaldococcus jannaschii |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Km at 50 mM NaCl not measurable | Methanocaldococcus jannaschii | |
0.012 | - |
polyA*dT36 | pH 8.0, 500 mM NaCl, wild-type enzyme | Methanocaldococcus jannaschii |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | 1:1 binding stoichiometry in absence of substrate at pH 8.0, activates, no binding to the enzyme but still weak activation without substrate at pH 6.5 | Methanocaldococcus jannaschii | |
Mn2+ | 1:1 binding stoichiometry in absence of substrate at pH 8.0, best activator, maximal activity at 10 mM and pH 8.0 | Methanocaldococcus jannaschii | |
additional information | enzyme is divalent metal ion-dependent, one metal ion binding mechanism, pH-dependence, kinetics, and thermodynamics for Mg2+, Mn2+, wild-type and mutant enzymes, substrate is involved in metal ion positioning and binding, Ca2+ and Ba2+ cannot substitute for Mn2+ or Mg2+ | Methanocaldococcus jannaschii |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Methanocaldococcus jannaschii | - |
type 2 enzyme | - |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli, partially | Methanocaldococcus jannaschii |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
Endonucleolytic cleavage to a 5'-phosphomonoester | Asp149 is essential for catalytic activity, Asp7, Glu8 and Asp112 are invovled in metal ion binding | Methanocaldococcus jannaschii |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | no activity with dT36 and polyA | Methanocaldococcus jannaschii | ? | - |
? | |
polyA*dT36 + H2O | - |
Methanocaldococcus jannaschii | ? | - |
? | |
polyA*dT36 hybrid + H2O | - |
Methanocaldococcus jannaschii | ? | - |
? | |
RNA*DNA hybrid + H2O | specifically degrades the RNA moiety | Methanocaldococcus jannaschii | 5'-phospho-3'-hydroxyoligonucleotides | - |
? |
Synonyms | Comment | Organism |
---|---|---|
RNase H | - |
Methanocaldococcus jannaschii |
RNase HII | - |
Methanocaldococcus jannaschii |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.11 | - |
polyA*dT36 | pH 8.0, 500 mM NaCl, wild-type enzyme | Methanocaldococcus jannaschii | |
0.64 | - |
polyA*dT36 | pH 8.0, 50 mM NaCl, wild-type enzyme | Methanocaldococcus jannaschii |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
at 10 mM Mn2+ or Mg2+, wild-type enzyme | Methanocaldococcus jannaschii |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6.5 | 8 | in presence of 10 mM Mn2+ the wild-type enzyme shows maximal activity at pH 8.0, 58% activity at pH 6.5, no activity at pH 5.5 | Methanocaldococcus jannaschii |