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Literature summary for 2.7.7.7 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
Search for more literature for 2.7.7.7

Cloned(Commentary)

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

Protein Variants

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

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

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ PolX has Mg2+-dependent 5'-3' DNA polymerase activity Thermus thermophilus
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]

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

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
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 ?
-
 ?
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

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

Purification (Commentary)

Purification (Comment) Organism
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)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
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
-
 ?
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
-
 ?
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 ?
-
 ?
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

Synonyms Comment Organism
DNA polymerase X
-
 Thermus thermophilus
Pol I
-
 Thermus thermophilus

kcat/KM [mM/s]

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