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Literature summary for 4.6.1.24 extracted from

  • Kobayashi, H.; Motoyoshi, N.; Itagaki, T.; Suzuki, M.; Inokuchi, N.
    Effect of the replacement of aspartic acid/glutamic acid residues with asparagine/glutamine residues in RNase He1 from Hericium erinaceus on inhibition of human leukemia cell line proliferation (2015), Biosci. Biotechnol. Biochem., 79, 211-217.
    View publication on PubMed

Cloned(Commentary)

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

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

Organism UniProt Comment Textmining
Hericium erinaceus B1Q4V2
-
-
Pleurotus ostreatus P81762
-
-

Purification (Commentary)

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

Source Tissue Comment Organism Textmining

Substrates and Products (Substrate)

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

Synonyms Comment Organism
RNase He1
-
Hericium erinaceus
RNase Po1
-
Pleurotus ostreatus
RNase T1
-
Hericium erinaceus
RNase T1
-
Pleurotus ostreatus

Temperature Optimum [°C]

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

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

pI Value

Organism Comment pI Value Maximum pI Value
Hericium erinaceus
-
-
4.2
Pleurotus ostreatus
-
-
9

General Information

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