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Literature summary for 2.7.7.8 extracted from

  • Wong, A.G.; McBurney, K.L.; Thompson, K.J.; Stickney, L.M.; Mackie, G.A.
    The S1 and KH domains of polynucleotide phosphorylase determine the efficiency of RNA binding and autoregulation (2013), J. Bacteriol., 195, 2021-2031.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.7.7.8

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant enzymes in Escherichia coli strain ENS134-3 from modofied plasmid pAW101 and in beta-galactosidase reporter strain IBPC7322(lambdaGF2), overview Escherichia coli

Protein Variants

Protein Variants Comment Organism
F635A site-directed mutagenesis, the mutant enzyme shows reduced activity compared to the wild-type enzyme Escherichia coli
F635A/F638A/H650A site-directed mutagenesis, the mutant enzyme shows highly reduced activity and an increased RNA binding constant compared to the wild-type enzyme Escherichia coli
F635R/F638R/H650R site-directed mutagenesis, the mutant enzyme shows reduced activity and an increased RNA binding constant compared to the wild-type enzyme Escherichia coli
F638A site-directed mutagenesis, the mutant enzyme shows reduced activity and an increased RNA binding constant compared to the wild-type enzyme Escherichia coli
G570C site-directed mutagenesis, the mutant enzyme shows highly reduced activity and an increased RNA binding constant compared to the wild-type enzyme Escherichia coli
G570C/V679A site-directed mutagenesis, the mutant enzyme shows reduced activity compared to the wild-type enzyme Escherichia coli
H650A site-directed mutagenesis, the mutant enzyme shows reduced activity compared to the wild-type enzyme Escherichia coli
I555T site-directed mutagenesis, the mutant enzyme shows slightly reduced activity compared to the wild-type enzyme Escherichia coli
I576A site-directed mutagenesis, the mutant enzyme shows reduced activity and an increased RNA binding constant compared to the wild-type enzyme Escherichia coli
I576A/F638A site-directed mutagenesis, the mutant enzyme shows reduced activity compared to the wild-type enzyme Escherichia coli
I576T site-directed mutagenesis, the mutant enzyme shows reduced activity and an increased RNA binding constant compared to the wild-type enzyme Escherichia coli
I576T/F638A site-directed mutagenesis, the mutant enzyme shows reduced activity and an increased RNA binding constant compared to the wild-type enzyme Escherichia coli
I576T/T585A site-directed mutagenesis, the mutant enzyme shows reduced activity compared to the wild-type enzyme Escherichia coli
K571L site-directed mutagenesis, the mutant enzyme shows reduced activity and an increased RNA binding constant compared to the wild-type enzyme Escherichia coli
K571Q site-directed mutagenesis, the mutant enzyme shows reduced activity and an increased RNA binding constant compared to the wild-type enzyme Escherichia coli
additional information construction of domain deletion mutants DELTAKH DELTAS1, DELTAKH, and DELTAS1 Escherichia coli

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Escherichia coli

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes from Escherichia coli strain ENS134-3 Escherichia coli

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
 recombinant enzyme activity is measured as repression of beta-galactosidase activity in the reporter expression sytem, RNA binding affinity and in vitro RNA-binding activities of wild-type and mutant enzymes, overview Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
RNAn+1 + phosphate substrate is synthetic radiolabeled SL9A RNA Escherichia coli RNAn + a nucleoside diphosphate
-
 ?

Subunits

Subunits Comment Organism
monomer domain organization of the enzyme monomer, homology modeling with KH domain and S1 domain, overview Escherichia coli

Synonyms

Synonyms Comment Organism
PNPase
-
 Escherichia coli
polynucleotide phosphorylase
-
 Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Escherichia coli

General Information

General Information Comment Organism
evolution polynucleotide phosphorylase is a conserved, widely distributed phosphorolytic 3'-5' exoribonuclease Escherichia coli
additional information the S1 and KH domains of polynucleotide phosphorylase determine the efficiency of RNA binding and enzyme autoregulation, modeling of the roles of the KH and S1 domains in PNPase-RNA interactions and in substrate binding, overview Escherichia coli