Cloned (Comment) | Organism |
---|---|
recombinant enzyme expression in Escherichia coli strain BL21(DE3) pLysS | Pleurotus ostreatus |
Crystallization (Comment) | Organism |
---|---|
purified recombinant enzyme, hanging drop vapor diffusion method, mixing of 500 nl of 10 mg/mL protein in 20 mM Tris-HCl, pH 7.5, with 500 nl of reservoir solution containing 4 M sodium formate, 0.1 M Bis-Tris propane, pH 7.0, 10% PEG 400, and 300 nl 2 M caesium chloride, 20°C, 3 days, X-ray diffraction structure determination and analysis at 1.85 A resolution, molecular replacement method | Pleurotus ostreatus |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
additional information | RNase Po1 might bind to the plasma membrane electrostatically | Pleurotus ostreatus | - |
- |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pleurotus ostreatus | P81762 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant enzyme from Escherichia coli strain BL21(DE3)pLysS to homogeneity by ammonium sulfate fractionation, gel filtration, cation exchange and anion exchange chromatography, and again gel filtration, heparin affinity chromatography, and dialysis | Pleurotus ostreatus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | the enzyme is a guanylic acid-specific ribonuclease | Pleurotus ostreatus | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | comparison of primary, secondary, and tertiary structures of RNase Po1 and RNase T1, overview | Pleurotus ostreatus |
Synonyms | Comment | Organism |
---|---|---|
RNase Po1 | - |
Pleurotus ostreatus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Pleurotus ostreatus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
- |
Pleurotus ostreatus |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the RNase T1 family, comparison of the amino acid sequences, overview | Pleurotus ostreatus |
additional information | comparison of the electrostatic potential of the molecular surfaces of RNase Po1 and RNase T1 shows that RNase T1 is anionic whereas RNase Po1 is cationic, so RNase Po1 might bind to the plasma membrane electrostatically, determination of the three-dimensional X-ray structure of RNase Po1 and comparison to that of RNase T1. One of the additional disulfide bond is in the catalytic and binding site of RNase Po1, and makes RNase Po1 more stable than RNase T1. The base recognition site of RNase T1 consists of Tyr42, Asn43, Asn44, Glu46, Tyr45, and Asn98 and is located in the loop between beta3-4 strands (Asn43, Asn44, Tyr45, Glu46) and in the loop between beta6-7 strands (Asn98). In case of the base recognition site of RNase Po1, the amino acid residues Tyr38, Asn39, Asn40, Phe41, Glu42, and Asn94 correspond to those of RNase T1 | Pleurotus ostreatus |
physiological function | the enzyme is cytotoxic and inhibits the proliferation of human tumor cells, the enzyme is internalized into tumour cells | Pleurotus ostreatus |