Cloned (Comment) | Organism |
---|---|
expression of the His-tagged N-terminal RNaseE catalytic N domain, comprising residues 1-529, in strain BL21(DE3) | Escherichia coli |
Crystallization (Comment) | Organism |
---|---|
purified recombinant His-tagged N-terminal RNaseE catalytic N domain, vapour diffusion method, 7 mg/ml protein in solution is mixed in a 1:1 ratio with precipitation solution containing 0.18 M Li2SO4, 0.09 M Tris-HCl, pH 8.5, 27% w/v PEG 4000, and 10% v/v glycerol, X-ray diffraction structure determination and analysis at 3.4 A resolution | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Escherichia coli |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
300000 | - |
N-terminal catalytic domain homotetramer, analytical ultracentrifugation | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged N-terminal RNaseE catalytic N domain from strain BL21(DE3) by metal affinity chromatography and gel filtration to homogeneity | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
5'ACAGUAUUUG-fluorescein + H2O | 5' monophosphorylated, 3' fluorescein-labeled synthetic substrate with protective 2'-O-methyl groups at all positions based on the 5' cleavage site of pBR322 RNA I | Escherichia coli | 5'ACAGU + AUUUG-fluorescein | - |
? | |
additional information | RNase E acts via a scanning mechanism in processing and degradation of RNA | Escherichia coli | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | the enzymes' conserved N-terminal catalytic domain forms homotetramers binding up to four molecules of specific RNA substrate, the tetramers forms a D2 dihedral symmetry, X-ray scattering, analytical ultracentrifugation, mass spectrometry, and circular dichroism used for structure analysis, overview | Escherichia coli |
Synonyms | Comment | Organism |
---|---|---|
ribonuclease E | - |
Escherichia coli |
RNase E | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.3 | - |
assay at | Escherichia coli |