Cloned (Comment) | Organism |
---|---|
recombinant expression of the His-tagged enzyme in Escherichia coli strain BL21(AI) | Escherichia coli |
Crystallization (Comment) | Organism |
---|---|
purified enzyme in complex with four different ssDNA substrates, 5'-Cy5-dT13, 5'-Cy5-dA13, dA16 and dT13, hanging drop vapor diffusion, 0.002 ml of 10 mg/ml enzyme in 20 mM Tris, 1 mM DTT, 10 mM EDTA, pH 8.0, with a 1.2 molar excess of oligonucleotide, is mixed with 0.002 ml of reservoir solution containing 0.9-1.5M ammonium sulfate, 3.756.0% 2-propanol and 25% glycerol, 1 week, X-ray diffraction structure determination and analysis at 2.0-3.7 A resolution | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required, binds at the active site, the Mg2+ ion is coordinated with near octahedral geometry to the scissile phosphate, the carboxylate of Asp15 and three water molecules | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P04995 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli strain BL21(AI) by nickel affinity chromatography and ultrafiltration | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | exonuclease I digests single-stranded DNA in the 3'-5' direction in a highly processive manner. The interactions at the anchor site, which involve all three domains of the enzyme protein and three consecutive nucleotides of the ssDNA | Escherichia coli | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
Exo1 | - |
Escherichia coli |
exonuclease 1 | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.5 | - |
assay at | Escherichia coli |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme is a member of the RAD2 nuclease family | Escherichia coli |
additional information | the enzyme crystal structure, in the absence of DNA, shows a C-shaped molecule with three domains that form a central positively charged groove. The active site is at the bottom of the groove, while an extended loop, proposed to encircle the DNA, crosses over the groove. Analysis of the active site structure of enzyme with bound ssDNA nucleotides, overview | Escherichia coli |