Cloned (Comment) | Organism |
---|---|
DNA and amino acid sequence determination and analysis, sequence comparisons, expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) pLysS | Hericium erinaceus |
Protein Variants | Comment | Organism |
---|---|---|
D19N/D22N/E25Q/D31N/D38N/E50Q/E57Q/E76Q/D77N/D79N/E92Q/D93N | site-directed mutagenesis using three different PCR primers | Hericium erinaceus |
D31N/D38N/E92Q/D93N | site-directed mutagenesis using two different PCR primers | Hericium erinaceus |
additional information | generation of enzyme mutants by replacing 12 Asn/Gln residues with Asp/Glu residues in analogy to the amino acid sequence of RNase Po1 from Pleurotus ostreatus. One mutant shows modified higher optimal pH for enzyme activity compared to the wild-type enzyme and inhibits the proliferation of cells in a human leukemia cell line | Hericium erinaceus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Hericium erinaceus | B1Q4V2 | - |
- |
Pleurotus ostreatus | P81762 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes 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 | Hericium erinaceus |
Source Tissue | Comment | Organism | Textmining |
---|
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | assay substrate is commercial yeast RNA, rates of release of GMP and cGMP show base specificity of enzyme mutant D19N/D22N/E25Q/D31N/D38N/E50Q/E57Q/E76Q/D77N/D79N/E92Q/D93N | Hericium erinaceus | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
RNase He1 | - |
Hericium erinaceus |
RNase Po1 | - |
Pleurotus ostreatus |
RNase T1 | - |
Hericium erinaceus |
RNase T1 | - |
Pleurotus ostreatus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
65 | 70 | - |
Pleurotus ostreatus |
65 | 70 | wild-type enzyme and recombinant mutant D19N/D22N/E25Q/D31N/D38N/E50Q/E57Q/E76Q/D77N/D79N/E92Q/D93N | Hericium erinaceus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
4.5 | - |
wild-type enzyme | Hericium erinaceus |
7 | - |
recombinant mutant D31N/D38N/E92Q/D93N | Hericium erinaceus |
7.5 | - |
- |
Pleurotus ostreatus |
7.5 | - |
recombinant mutant D19N/D22N/E25Q/D31N/D38N/E50Q/E57Q/E76Q/D77N/D79N/E92Q/D93N | Hericium erinaceus |
Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|
Hericium erinaceus | - |
- |
4.2 |
Pleurotus ostreatus | - |
- |
9 |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme is a member of the RNase T1 family | Hericium erinaceus |
evolution | the enzyme is a member of the RNase T1 family | Pleurotus ostreatus |
physiological function | the enzyme inhibits human tumor cell line proliferation | Pleurotus ostreatus |
physiological function | the wild-type enzyme shows little inhibition of human tumor cell proliferation, but the mutant D19N/D22N/E25Q/D31N/D38N/E50Q/E57Q/E76Q/D77N/D79N/E92Q/D93N is inhibiting proliferation in human leukemia cell lines, HL-60 and Jurkat with IC50 values of 100 nM and 0.002 mM, respectively, mutant D31N/D38N/E92Q/D93N is inactive | Hericium erinaceus |