Literature summary for 3.1.13.1 extracted from

Reis, F.P.; Barria, C.; Gomez-Puertas, P.; Gomes, C.M.; Arraiano, C.M.
Identification of temperature-sensitive mutations and characterization of thermolabile RNase II variants (2019), FEBS Lett., 593, 352-360 .

Search for more literature for 3.1.13.1

Cloned(Commentary)

Cloned (Comment)	Organism
recombinant expression of His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3), enzyme expression for complementation in Escherichia coli MG1693-derivative strain CMA401	Escherichia coli

Protein Variants

Protein Variants	Comment	Organism
C284Y	RNase II thermolability of the rnb500 phenotype is due to the Cys284Tyr mutation within the RNB domain, which abolishes activity by increasing protein kinetic instability at the nonpermissive temperature. Expression of RNase II C284Y and the double mutant (D126N and C284Y) does not allow growth at the nonpermissive temperature of 44°C. Structural mapping and partial multiple sequence alignment of RNase II thermosensitive phenotype mutations, overview	Escherichia coli
D126N	site-directed mutagenesis, the RNase II mutant exhibits a slightly decreased growth at 44°C suggesting some thermosensitivity which does not account for a major phenotype. This individual mutation is not detrimental for the function of RNase II in vivo. the RNase II D126N variant exhibits substantial catalytic activity	Escherichia coli
D126N/C284Y	site-directed mutagenesis, expression of RNase II C284Y and the double mutant (D126N and C284Y) does not allow growth at the nonpermissive temperature of 44°C	Escherichia coli

Organism

Organism	UniProt	Comment	Textmining
Escherichia coli	P30850	-	-
Escherichia coli SK5689	P30850	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography	Escherichia coli

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
additional information	the enzyme substrate is a 30mer oligoribonucleotide (5' CCCGACACCAACCACUAAAAAAAAAAAAAA-3') labeled at 5'-end with [c-32P]ATP through T4 polynucleotide kinase	Escherichia coli	?	-	?
additional information	the enzyme substrate is a 30mer oligoribonucleotide (5' CCCGACACCAACCACUAAAAAAAAAAAAAA-3') labeled at 5'-end with [c-32P]ATP through T4 polynucleotide kinase	Escherichia coli SK5689	?	-	?

Synonyms

Synonyms	Comment	Organism
RNase II	-	Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
30	-	assay at	Escherichia coli

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
44	-	enzyme inactivation, RNase II is thermolabile	Escherichia coli

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8	-	assay at	Escherichia coli

General Information

General Information	Comment	Organism
malfunction	RNase II thermolability of the rnb500 phenotype is due to the Cys284Tyr mutation within the RNB domain, which abolishes activity by increasing protein kinetic instability at the nonpermissive temperature. In vivo and in vitro investigation of RNase II mutation(s) that confer the rnb500 phenotype	Escherichia coli

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Release 2023.1 (January 2023)