Cloned (Comment) | Organism |
---|---|
overexpression of wild-type and several truncated enzymes in Escherichia coli | Thermococcus kodakarensis |
Crystallization (Comment) | Organism |
---|---|
purified truncated enzyme RNase HII-213, comprising residues 1-213, metal-bound or metal-free, hanging drop vapour diffusion method, 15% w/v PEG 6000, 200 mM MES, pH 6.5, X-ray diffraction structure determinaion and analysis at 3.5 A resolution | Thermococcus kodakarensis |
Protein Variants | Comment | Organism |
---|---|---|
D105A | site-directed mutagenesis, nearly inactive mutant | Thermococcus kodakarensis |
D135A | site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme | Thermococcus kodakarensis |
D7A | site-directed mutagenesis, nearly inactive mutant | Thermococcus kodakarensis |
E8A | site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme | Thermococcus kodakarensis |
H132A | site-directed mutagenesis, reduced activity compared to the wild-type enzyme | Thermococcus kodakarensis |
additional information | construction of several truncation mutants, comprising residues 1-198, 1-203, 1-207, 1-213, or 1-217 of the wild-type enzyme, the truncated mutants' activities are increased or reduced compared to the wild-type enzyme, overview | Thermococcus kodakarensis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00009 | - |
RNA*DNA hybrid | wild-type enzyme, pH 8.0, 30°C | Thermococcus kodakarensis | |
0.00054 | - |
RNA*DNA hybrid | mutant D135A, pH 8.0, 30°C | Thermococcus kodakarensis | |
0.00097 | - |
RNA*DNA hybrid | truncated mutant RNase HII-217, pH 8.0, 30°C | Thermococcus kodakarensis | |
0.00118 | - |
RNA*DNA hybrid | mutant E8A, pH 8.0, 30°C | Thermococcus kodakarensis | |
0.0014 | - |
RNA*DNA hybrid | mutant H132A, pH 8.0, 30°C | Thermococcus kodakarensis | |
0.00155 | - |
RNA*DNA hybrid | truncated mutant RNase HII-207, pH 8.0, 30°C | Thermococcus kodakarensis | |
0.0019 | - |
RNA*DNA hybrid | truncated mutant RNase HII-213, pH 8.0, 30°C | Thermococcus kodakarensis | |
0.00837 | - |
RNA*DNA hybrid | truncated mutant RNase HII-203, pH 8.0, 30°C | Thermococcus kodakarensis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Co2+ | - |
Thermococcus kodakarensis | |
Mg2+ | - |
Thermococcus kodakarensis | |
Mn2+ | - |
Thermococcus kodakarensis | |
additional information | type 2 enzyme requires divalent cations | Thermococcus kodakarensis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermococcus kodakarensis | - |
type 2 enzyme | - |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and truncated enzymes from Escherichia coli | Thermococcus kodakarensis |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
Endonucleolytic cleavage to a 5'-phosphomonoester | the catalytic reaction requires 4 acidic residues, catalytic center and catalytic mechanism of type 1 and type 2 ribonucleases, type 1 enzyme requires a histidine as general base, while type 2 enzyme does not, substrate binding of type 2 enzyme | Thermococcus kodakarensis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
RNA*DNA hybrid + H2O | the C-terminal domain of type 2 enzyme is involved in the interaction with the substrate | Thermococcus kodakarensis | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
KOD1 | - |
Thermococcus kodakarensis |
ribonuclease H | - |
Thermococcus kodakarensis |
RNase H | - |
Thermococcus kodakarensis |
Tk-RNase HII | - |
Thermococcus kodakarensis |
type 2 ribonuclease H | - |
Thermococcus kodakarensis |
type 2 RNase H | - |
Thermococcus kodakarensis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Thermococcus kodakarensis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Thermococcus kodakarensis |