Literature summary for 3.1.21.1 extracted from
Ho, T.Y.; Wu, S.L.; Hsiang, C.H.; Chang, T.J.; Hsiang, C.Y.
Identification of a DNA-binding domain and an active-site residue of pseudorabies virus DNase (2000), Biochem. J., 346, 441-445.
No PubMed abstract available
Cloned(Commentary)
Cloned (Comment) |
Organism |
expression of wild-type and deletion mutants in Escherichia coli BL21 (DE3) |
Suid alphaherpesvirus 1 |
Protein Variants
Protein Variants |
Comment |
Organism |
H371A |
site-directed mutagenesis, exchange of a well-conserved His residue, loss of most of the nuclease activity, stilla ctive in DNA-binding |
Suid alphaherpesvirus 1 |
additional information |
construction of mutants with deletions at the N- and/or the C-terminus, mutants reveal the localisation fo activities within the enzyme sequence, overview |
Suid alphaherpesvirus 1 |
Metals/Ions
Metals/Ions |
Comment |
Organism |
Structure |
Mg2+ |
- |
Suid alphaherpesvirus 1 |
|
Organism
Organism |
UniProt |
Comment |
Textmining |
Suid alphaherpesvirus 1 |
- |
i.e. PRV |
- |
Purification (Commentary)
Purification (Comment) |
Organism |
recombinant wild-type and deletion mutants from Escherichia coli, to homogeneity |
Suid alphaherpesvirus 1 |
Reaction
Reaction |
Comment |
Organism |
Reaction ID |
endonucleolytic cleavage to 5'-phosphodinucleotide and 5'-phosphooligonucleotide end-products |
catalytic mechanism, the N-terminal 62 residues and the C-terminal 39 residues have important roles in 3'-exonuclease activity, residues 63-453 are responsible for 5'- and 3'-exonuclease activities, residues 24-492 form a region with 7fold higher catalytic activity than the intact enzyme |
Suid alphaherpesvirus 1 |
|
Specific Activity [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg] |
Specific Activity Maximum [µmol/min/mg] |
Comment |
Organism |
additional information |
- |
5'-exonuclease, 3'-exonuclease and DNA-binding activities of wild-type and truncated mutants and mutant H371A |
Suid alphaherpesvirus 1 |
Substrates and Products (Substrate)
Substrates |
Comment Substrates |
Organism |
Products |
Comment (Products) |
Rev. |
Reac. |
pUC18 DNA + H2O |
- |
Suid alphaherpesvirus 1 |
? |
- |
? |
|
Subunits
Subunits |
Comment |
Organism |
More |
the enzyme is divided into a catalytic domain and a DNA-binding domain |
Suid alphaherpesvirus 1 |
Synonyms
Synonyms |
Comment |
Organism |
alkaline nuclease |
- |
Suid alphaherpesvirus 1 |
DNase |
- |
Suid alphaherpesvirus 1 |
Temperature Optimum [°C]
Temperature Optimum [°C] |
Temperature Optimum Maximum [°C] |
Comment |
Organism |
37 |
- |
assay at |
Suid alphaherpesvirus 1 |
pH Optimum
pH Optimum Minimum |
pH Optimum Maximum |
Comment |
Organism |
9 |
- |
assay at |
Suid alphaherpesvirus 1 |