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Literature summary for 3.1.26.4 extracted from

  • Goedken, E.R.; Keck, J.L.; Berger, J.M.; Marqusee, S.
    Divalent metal cofactor binding in the kinetic folding trajectory of Escherichia coli ribonuclease HI (2000), Protein Sci., 9, 1914-1921.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
overexpression of wild-type and mutant D10A enzymes in Escherichia coli Escherichia coli

Crystallization (Commentary)

Crystallization (Comment) Organism
4-10 mg/ml purifed recombinant wild-type and mutant D10A enzymes, in 20 mM HEPES, pH 7.0-8.0, 5-15% PEG 3350, hanging drop vapour diffusion method and microseeding, several weeks, X-ray diffraction structure determination and analysis Escherichia coli

Protein Variants

Protein Variants Comment Organism
D10A site-directed mutagenesis, active site mutant, no metal binding, altered folding kinetics in absence of metal ions to the values for wild-type enzyme in presence of metal ions Escherichia coli

General Stability

General Stability Organism
metal binding stabilizes the enzyme by decreasing its unfolding rate, mechanism Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information folding kinetics of the enzyme with Mg2+ bound or metal-free Escherichia coli

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ binding involves Asp10 and is pH-dependent, binds in the active site pocket of the natively folded enzyme only, stabilizes the enzyme conformation, effect of metal binding on enzyme folding kinetics Escherichia coli

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
-
-

Reaction

Reaction Comment Organism Reaction ID
Endonucleolytic cleavage to a 5'-phosphomonoester Asp10 is critical for activity and involved in binding of divalent metal ion Escherichia coli

Subunits

Subunits Comment Organism
More enzyme folding pathway Escherichia coli

Synonyms

Synonyms Comment Organism
ribonuclease H
-
Escherichia coli
ribonuclease HI
-
Escherichia coli
RNase H
-
Escherichia coli
RNase HI
-
Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
additional information
-
folding/unfolding and metal binding at different pH values Escherichia coli