Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Saccharolobus solfataricus |
Crystallization (Comment) | Organism |
---|---|
the enzyme is mixed with DNA (1:1.2 molar ratio) in 20 mM Tris-HCl buffer (pH 8.0, 25 °C) containing 60 mM NaCl, 4% glycerol (v/v), and 5 mM 2-mercaptoethanol and then placed on ice for 1 h prior to incubation with 5 mM MgCl2 and 1 mM dGTP. Crystals are grown using the sitting drop/vapor-diffusion method with the reservoir solution containing 20 mM Tris-HCl (pH 8.0 at 25 °C), 15% polyethylene glycol 3350 (w/v), 60 mM NaCl, 5 mM MgCl2, and 4% glycerol (v/v). Two crystal structures of Dpo4 with a template N2-NaphG (in a post-insertion register opposite a 3-terminal C in the primer) are solved. One shows N2-NaphG in a syn conformation, with the naphthyl group located between the template and the Dpo4 little finger domain. The Hoogsteen face is within hydrogen bonding distance of the N4 atoms of the cytosine opposite N2-NaphG and the cytosine at the 2 position. The second structure shows N2-NaphG in an anti conformation with the primer terminus largely disordered | Saccharolobus solfataricus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | steady-state kinetic parameters for one-base incorporation opposite G and N2-alkyl G adducts by Dpo4. Steady-state kinetic parameters for next base extension from G (or N2-alkyl G):C (or T) template primer termini by Dpo4 | Saccharolobus solfataricus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
deoxynucleoside triphosphate + DNAn | Saccharolobus solfataricus | - |
diphosphate + DNAn+1 | - |
? | |
deoxynucleoside triphosphate + DNAn | Saccharolobus solfataricus P2 | - |
diphosphate + DNAn+1 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Saccharolobus solfataricus | Q97W02 | - |
- |
Saccharolobus solfataricus P2 | Q97W02 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
deoxynucleoside triphosphate + DNAn | - |
Saccharolobus solfataricus | diphosphate + DNAn+1 | - |
? | |
deoxynucleoside triphosphate + DNAn | the enzyme shows a limited decrease in catalytic efficiency (kcat/Km) for insertion of dCTP opposite a series of N2-alkylguanine templates of increasing size from (methyl (Me) to (9-anthracenyl)-Me (Anth)). Fidelity is maintained with increasing size up to (2-naphthyl)-Me (Naph). The catalytic efficiency increases slightly going from the N2-NaphG to the N2-AnthG substrate, at the cost of fidelity. A set of oligonucleotides differing only in their N2-substitution at a single G site is used in this study with Dpo4 | Saccharolobus solfataricus | diphosphate + DNAn+1 | - |
? | |
deoxynucleoside triphosphate + DNAn | - |
Saccharolobus solfataricus P2 | diphosphate + DNAn+1 | - |
? | |
deoxynucleoside triphosphate + DNAn | the enzyme shows a limited decrease in catalytic efficiency (kcat/Km) for insertion of dCTP opposite a series of N2-alkylguanine templates of increasing size from (methyl (Me) to (9-anthracenyl)-Me (Anth)). Fidelity is maintained with increasing size up to (2-naphthyl)-Me (Naph). The catalytic efficiency increases slightly going from the N2-NaphG to the N2-AnthG substrate, at the cost of fidelity. A set of oligonucleotides differing only in their N2-substitution at a single G site is used in this study with Dpo4 | Saccharolobus solfataricus P2 | diphosphate + DNAn+1 | - |
? |
Synonyms | Comment | Organism |
---|---|---|
DNA polymerase Dpo4 | - |
Saccharolobus solfataricus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | steady-state kinetic parameters for one-base incorporation opposite G and N2-alkyl G adducts by Dpo4. Steady-state kinetic parameters for next base extension from G (or N2-alkyl G):C (or T) template primer termini by Dpo4 | Saccharolobus solfataricus |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | steady-state kinetic parameters for one-base incorporation opposite G and N2-alkyl G adducts by Dpo4. Steady-state kinetic parameters for next base extension from G (or N2-alkyl G):C (or T) template primer termini by Dpo4 | Saccharolobus solfataricus |