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Literature summary extracted from

  • Nakane, S.; Nakagawa, N.; Kuramitsu, S.; Masui, R.
    Characterization of DNA polymerase X from Thermus thermophilus HB8 reveals the POLXc and PHP domains are both required for 3-5 exonuclease activity (2009), Nucleic Acids Res., 37, 2037-2052.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.7.7 expressed in Escherichia coli Rosetta(DE3) cells Thermus thermophilus

Protein Variants

EC Number Protein Variants Comment Organism
2.7.7.7 D349A the Mg2+-dependent DNA polymerase activity of the mutant is almost the same as that of wild type enzyme Thermus thermophilus
2.7.7.7 D529A the Mg2+-dependent DNA polymerase activity of the mutant is almost the same as that of wild type enzyme Thermus thermophilus
2.7.7.7 E413A the Mg2+-dependent DNA polymerase activity of the mutant is almost the same as that of wild type enzyme Thermus thermophilus
2.7.7.7 H344A the Mg2+-dependent DNA polymerase activity of the mutant is almost the same as that of wild type enzyme Thermus thermophilus
2.7.7.7 H374A the Mg2+-dependent DNA polymerase activity of the mutant is almost the same as that of wild type enzyme Thermus thermophilus
2.7.7.7 H440A the Mg2+-dependent DNA polymerase activity of the mutant is almost the same as that of wild type enzyme Thermus thermophilus
2.7.7.7 H468A the Mg2+-dependent DNA polymerase activity of the mutant is almost the same as that of wild type enzyme Thermus thermophilus
2.7.7.7 H531A the Mg2+-dependent DNA polymerase activity of the mutant is almost the same as that of wild type enzyme Thermus thermophilus
2.7.7.7 Q342A the Mg2+-dependent DNA polymerase activity of the mutant is almost the same as that of wild type enzyme Thermus thermophilus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.7.7 0.0013
-
 DNAn in the presence of dNTPs Thermus thermophilus
2.7.7.7 0.0017
-
 DNAn in the absence of dNTPs Thermus thermophilus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.7.7 Mg2+ PolX has Mg2+-dependent 5'-3' DNA polymerase activity Thermus thermophilus
2.7.7.7 additional information the PolX catalytic domain exhibits no polymerase activity with 5 mM Zn2+, Ni2+, Ca2+ or Co2+, or without a metal ion Thermus thermophilus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.7.7.7 64000
-
 estimated from SDS-PAGE Thermus thermophilus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.7.7 additional information Thermus thermophilus the catalytic efficiency of PolX is almost the same with and without dNTPs, whereas that of the domain mixture increases on the addition of dNTPs ?
-
 ?
2.7.7.7 additional information Thermus thermophilus HB8 / ATCC 27634 / DSM 579 the catalytic efficiency of PolX is almost the same with and without dNTPs, whereas that of the domain mixture increases on the addition of dNTPs ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.7.7.7 Thermus thermophilus
-
-
-
2.7.7.7 Thermus thermophilus HB8 / ATCC 27634 / DSM 579
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.7.7 ammonium sulfate precipitation, Toyopearl phenyl-650M column chroamtography, Super Q-650M column chromatography, hydroxyapatite column chromatography, Superdex 200 gel filtration, His-bind resin column chromatography, Toyopearl butyl column chromatography, and Superdex 75 gel filtration Thermus thermophilus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.7.7 deoxynucleoside triphosphate + DNAn in the presence of Mg2+ or Mn2+, the POLX catalytic domain inserts dIMP, IMP and 8-oxo-dGMP opposite deoxycytosine as well as dGMP and GMP, PolX likely prefers deoxyguanine to deoxythymidine Thermus thermophilus diphosphate + DNAn+1
-
 ?
2.7.7.7 deoxynucleoside triphosphate + DNAn in the presence of Mg2+ or Mn2+, the POLX catalytic domain inserts dIMP, IMP and 8-oxo-dGMP opposite deoxycytosine as well as dGMP and GMP, PolX likely prefers deoxyguanine to deoxythymidine Thermus thermophilus HB8 / ATCC 27634 / DSM 579 diphosphate + DNAn+1
-
 ?
2.7.7.7 additional information the catalytic efficiency of PolX is almost the same with and without dNTPs, whereas that of the domain mixture increases on the addition of dNTPs Thermus thermophilus ?
-
 ?
2.7.7.7 additional information the catalytic efficiency of PolX is almost the same with and without dNTPs, whereas that of the domain mixture increases on the addition of dNTPs Thermus thermophilus HB8 / ATCC 27634 / DSM 579 ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.7.7.7 DNA polymerase X
-
 Thermus thermophilus
2.7.7.7 Pol I
-
 Thermus thermophilus

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.7.7.7 28
-
 DNAn in the absence of dNTPs Thermus thermophilus
2.7.7.7 38
-
 DNAn in the presence of dNTPs Thermus thermophilus