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Literature summary extracted from

  • Jain, R.; Shuman, S.
    Characterization of a thermostable archaeal polynucleotide kinase homologous to human Clp1 (2009), RNA, 15, 923-931.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.1.78 into the vector pET28-His10Smt3 for expression in Escherichia coli BL21DE3 cells Pyrococcus horikoshii

Protein Variants

EC Number Protein Variants Comment Organism
2.7.1.78 D73A mutation abolishes activity Pyrococcus horikoshii
2.7.1.78 K49A mutation reduces activity to 1% of the wild-type value Pyrococcus horikoshii

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.1.78 0.016
-
 5'-OH DNA
-
 Pyrococcus horikoshii
2.7.1.78 0.021
-
 5'-phospho-DNA
-
 Pyrococcus horikoshii

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.1.78 Mg2+
-
 Pyrococcus horikoshii

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.7.1.78 41000
-
 determined by SDS-PAGE, tag-free Pyrococcus horikoshii

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.1.78 ATP + 5'-OH DNA Pyrococcus horikoshii
-
 ADP + 5'-phospho-DNA
-
 r
2.7.1.78 ATP + 5'-OH RNA Pyrococcus horikoshii preference for RNA substrates ADP + 5'-phospho-RNA
-
 r

Organism

EC Number Organism UniProt Comment Textmining
2.7.1.78 Pyrococcus horikoshii Q57936
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.1.78 on a Ni-NTA-agarose column, the His10Smt3 tag is removed with the Smt3-specific protease Ulp1 Pyrococcus horikoshii

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.1.78 ATP + 5'-OH DNA
-
 Pyrococcus horikoshii ADP + 5'-phospho-DNA
-
 r
2.7.1.78 ATP + 5'-OH RNA preference for RNA substrates Pyrococcus horikoshii ADP + 5'-phospho-RNA
-
 r

Synonyms

EC Number Synonyms Comment Organism
2.7.1.78 5'-OH polynucleotide kinase
-
 Pyrococcus horikoshii
2.7.1.78 Clp1-like protein
-
 Pyrococcus horikoshii
2.7.1.78 PhoClp1
-
 Pyrococcus horikoshii
2.7.1.78 polynucleotide kinase
-
 Pyrococcus horikoshii

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.1.78 55
-
 kinase assay Pyrococcus horikoshii

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
2.7.1.78 55 85
-
 Pyrococcus horikoshii

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.1.78 0.00083
-
 5'-phospho-DNA
-
 Pyrococcus horikoshii
2.7.1.78 0.077
-
 5'-OH DNA
-
 Pyrococcus horikoshii

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.1.78 5
-
 reverse reaction Pyrococcus horikoshii
2.7.1.78 6.5
-
 kinase assay Pyrococcus horikoshii
2.7.1.78 7.5
-
 kinase assay Pyrococcus horikoshii

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.7.1.78 4.5 9.5 DNA kinase activity Pyrococcus horikoshii
2.7.1.78 4.5 6.5 RNA kinase activity Pyrococcus horikoshii

Cofactor

EC Number Cofactor Comment Organism Structure
2.7.1.78 ATP
-
 Pyrococcus horikoshii