Cloned (Comment) | Organism |
---|---|
overexpression of wild-type and mutant D10A enzymes in Escherichia coli | Escherichia coli |
Crystallization (Comment) | Organism |
---|---|
4-10 mg/ml purifed recombinant wild-type and mutant D10A enzymes, in 20 mM HEPES, pH 7.0-8.0, 5-15% PEG 3350, hanging drop vapour diffusion method and microseeding, several weeks, X-ray diffraction structure determination and analysis | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
D10A | site-directed mutagenesis, active site mutant, no metal binding, altered folding kinetics in absence of metal ions to the values for wild-type enzyme in presence of metal ions | Escherichia coli |
General Stability | Organism |
---|---|
metal binding stabilizes the enzyme by decreasing its unfolding rate, mechanism | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | folding kinetics of the enzyme with Mg2+ bound or metal-free | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | binding involves Asp10 and is pH-dependent, binds in the active site pocket of the natively folded enzyme only, stabilizes the enzyme conformation, effect of metal binding on enzyme folding kinetics | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
Endonucleolytic cleavage to a 5'-phosphomonoester | Asp10 is critical for activity and involved in binding of divalent metal ion | Escherichia coli |
Subunits | Comment | Organism |
---|---|---|
More | enzyme folding pathway | Escherichia coli |
Synonyms | Comment | Organism |
---|---|---|
ribonuclease H | - |
Escherichia coli |
ribonuclease HI | - |
Escherichia coli |
RNase H | - |
Escherichia coli |
RNase HI | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
additional information | - |
folding/unfolding and metal binding at different pH values | Escherichia coli |