Cloned (Comment) | Organism |
---|---|
overexpression of wild-type and mutant enzymes in Escherichia coli strain HB101 | Thermus thermophilus |
overexpression of wild-type and mutant enzymes in strain HB101 | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
C13A/C63A/C133A/E135C | site-directed mutagenesis, 37% activity compared to the wild-type enzyme | Escherichia coli |
C13S/C63A/R135C | site-directed mutagenesis, 6.9% activity compared to the wild-type enzyme | Thermus thermophilus |
C63A | site-directed mutagenesis, 94% activity compared to the wild-type enzyme | Thermus thermophilus |
C63A/R135C | site-directed mutagenesis, 44% activity compared to the wild-type enzyme | Thermus thermophilus |
additional information | construction of DNA-linked mutant enzyme, by cross-linking of the 15mer DNA, which is complementary to the polypurine-tract sequence of human immunodeficiency virus-1 RNA, i.e. PPT-RNA, to the thiol group of Cys13 or Cys135 of the enzyme, mutants shows 0.69% activity compared to the wild-type enzyme | Thermus thermophilus |
additional information | construction of DNA-linked mutant enzyme, by cross-linking of the 15mer DNA, which is complementary to the polypurine-tract sequence of human immunodeficiency virus-1 RNA, i.e. PPT-RNA, to the thiol group of Cys135 of the enzyme, mutant shows 4.2% activity compared to the wild-type enzyme | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Thermus thermophilus | - |
- |
- |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
Endonucleolytic cleavage to a 5'-phosphomonoester | active site residues are Asp10, Gu48, Asp70, His124, Asp134 | Thermus thermophilus |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
activity of wild-type and mutant enzymes, determination by spectrophotometric assay, at different temperatures | Thermus thermophilus |
additional information | - |
activity of wild-type and mutant enzymes, determination by spectrophotometric assay, at different temperatures | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
PPT-RNA + H2O | single-stranded, the DNA-linked enzyme mutant shows highly reduced activity compared to the wild-type enzyme, specific cleavage of the 15mer DNA, which is complementary to the polypurine-tract sequence of human immunodeficiency virus-1 RNA | Thermus thermophilus | ? | - |
? | |
PPT-RNA + H2O | single-stranded, the DNA-linked enzyme mutant shows highly reduced activity compared to the wild-type enzyme, specific cleavage of the 15mer DNA, which is complementary to the polypurine-tract sequence of human immunodeficiency virus-1 RNA | Escherichia coli | ? | - |
? | |
RNA*DNA hybrid + H2O | specific for | Thermus thermophilus | ? | - |
? | |
RNA*DNA hybrid + H2O | specific for | Escherichia coli | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | three-dimensional structure of wild-type enzyme | Thermus thermophilus |
Synonyms | Comment | Organism |
---|---|---|
endoribonuclease H (calf thymus) | - |
Thermus thermophilus |
ERNH | - |
Escherichia coli |
hybrid nuclease | - |
Thermus thermophilus |
hybrid ribonuclease | - |
Thermus thermophilus |
hybridase | - |
Thermus thermophilus |
hybridase (ribonuclease H) | - |
Thermus thermophilus |
ribonuclease H | - |
Thermus thermophilus |
ribonuclease H | - |
Escherichia coli |
RNA*DNA hybrid ribonucleotiohydrolase | - |
Thermus thermophilus |
RNase H | - |
Thermus thermophilus |
RNase H | - |
Escherichia coli |
RNase HI | - |
Thermus thermophilus |
RNase HI | - |
Escherichia coli |
TRNH | - |
Thermus thermophilus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at, substrate RNA*DNA hybrid | Thermus thermophilus |
30 | - |
assay at, substrate RNA*DNA hybrid | Escherichia coli |
50 | - |
DNA-C135-linked mutant, substrate PPT-RNA | Escherichia coli |
65 | - |
assay at, substrate PPT-RNA | Escherichia coli |
65 | - |
wild-type enzyme, assay at, substrate PPT-RNA, DNA-C135-linked mutant enzyme shows optimal activity with this substrate at 65°C | Thermus thermophilus |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | - |
DNA-linked mutant, loss of 50% activity after 15 min | Escherichia coli |
60 | - |
DNA-linked mutant, complete inactivation after 15 min | Escherichia coli |
90 | - |
DNA-linked mutant is almost fully stable after 15 min | Thermus thermophilus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Escherichia coli |
9 | - |
assay at | Thermus thermophilus |