Literature summary for 3.1.26.4 extracted from

Tsunaka, Y.; Takano, K.; Matsumura, H.; Yamagata, Y.; Kanaya, S.
Identification of single Mn2+ binding sites required for activation of the mutant proteins of E. coli RNase HI at Glu48 and/or Asp134 by X-ray crystallography (2005), J. Mol. Biol., 345, 1171-1183.

Search for more literature for 3.1.26.4

Cloned(Commentary)

Cloned (Comment)	Organism
overexpression of mutant enzymes in strain HB101	Escherichia coli

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant enzyme mutants with Mn2+ bound at the active sites, sitting drop vapour diffusion method, 6.7-9.9 mg/ml protein in 20 mM HEPES-NaOH, pH 7.0, 5-10 mm MnCl2, 20-28% PEG 3350, and 10% glycerol, 3 weeks, X-ray diffraction structure determination and analysis at 2.2-2.3 A resolution	Escherichia coli

Protein Variants

Protein Variants	Comment	Organism
D134A/L87A	site-directed mutagenesis, crystal structure analysis with bound Mn2+	Escherichia coli
E48A/L87A	site-directed mutagenesis, crystal structure analysis with bound Mn2+	Escherichia coli
E48A/L87A/D134A	site-directed mutagenesis, crystal structure analysis with bound Mn2+	Escherichia coli

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mn2+	activates, two single binding sites: site 1 is formed by Glu48, Asp10, and Asp70, site 2 is formed by Asp10 and Asp134, Glu48 and Asp134 are absolutely required for enzyme activation, binding structure and one-to-two metal mechanism, overview	Escherichia coli

Organism

Organism	UniProt	Comment	Textmining
Escherichia coli	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant mutant enzymes from strain HB101 by heparin affinity chromatography	Escherichia coli

Reaction

Reaction	Comment	Organism	Reaction ID
Endonucleolytic cleavage to a 5'-phosphomonoester	active site residues D10, E48, D70, and D134 are involved in metal ion binding, overview	Escherichia coli	a

Synonyms

Synonyms	Comment	Organism
RNase HI	-	Escherichia coli

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Release 2023.1 (January 2023)