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Literature summary for 3.1.30.1 extracted from

  • Guan, C.; Kumar, S.; Kucera, R.; Ewel, A.
    Changing the enzymatic activity of T7 endonuclease by mutations at the beta-bridge site: alteration of substrate specificity profile and metal ion requirements by mutation distant from the catalytic domain (2004), Biochemistry, 43, 4313-4322.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Escherichia phage T7

Crystallization (Commentary)

Crystallization (Comment) Organism
-
Escherichia phage T7

Protein Variants

Protein Variants Comment Organism
additional information mutation in beta-bridge center that separates the catalytic domains, 20-50 times lower nonspecific nuclease activity in Mg2+ buffer Escherichia phage T7

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required for resolving of cruciform structures Escherichia phage T7
Mn2+ 20times lower activity in resolving of cruciform structures than with Mg2+ Escherichia phage T7

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
DNA + H2O Escherichia phage T7 T7 endonuclase I resolves four-way junctions by simultaeously introducing two nicks on the two continously stacked strands at sites 5' to the junction, also acts on a variety of DNA structures nicked DNA
-
?

Organism

Organism UniProt Comment Textmining
Escherichia phage T7
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant T7 endonuclease Escherichia phage T7

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
0.6-0.8 U/mg, 1 unit is defined as the amount of enzmyme that is needed to convert 0.001 mg of supercoiled pUC(AT) to either the linear or nicked form in a 0.02 ml reaction at 37°C in 30 min Escherichia phage T7

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
DNA + H2O T7 endonuclase I resolves four-way junctions by simultaeously introducing two nicks on the two continously stacked strands at sites 5' to the junction, also acts on a variety of DNA structures Escherichia phage T7 nicked DNA
-
?

Subunits

Subunits Comment Organism
homodimer
-
Escherichia phage T7

Synonyms

Synonyms Comment Organism
resolvase T7
-
Escherichia phage T7
T7 endo I
-
Escherichia phage T7

pI Value

Organism Comment pI Value Maximum pI Value
Escherichia phage T7 deduced from nucleotide sequence
-
9.5