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Literature summary extracted from

  • Das, U.; Wang, L.K.; Smith, P.; Shuman, S.
    Structural and biochemical analysis of the phosphate donor specificity of the polynucleotide kinase component of the bacterial pnkp-hen1 RNA repair system (2013), Biochemistry, 52, 4734-4743.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.1.78 expressed in Escherichia coli BL21(DE3) cells Acetivibrio thermocellus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.7.1.78 in complexes with GTP, CTP, UTP and dATP, hanging drop vapor diffusion method, using 100 mM sodium citrate, pH 5.6, 100 mM MgCl2, 16-18% (w/v) PEG-3350 Acetivibrio thermocellus

Protein Variants

EC Number Protein Variants Comment Organism
2.7.1.78 R116A the mutation elicits a 10fold increase in Km for ATP, but has little effect on the turnover number value leading to 83% of wild type activity Acetivibrio thermocellus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.1.78 0.0028
-
 5'-HO-CCGACCAACGAAGGT wild type enzyme, at pH 7.0 and 37°C Acetivibrio thermocellus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.1.78 Mg2+ required Acetivibrio thermocellus

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'-dephospho-DNA Acetivibrio thermocellus
-
 ADP + 5'-phospho-DNA
-
 r
2.7.1.78 ATP + 5'-dephospho-RNA Acetivibrio thermocellus
-
 ADP + 5'-phospho-RNA
-
 r

Organism

EC Number Organism UniProt Comment Textmining
2.7.1.78 Acetivibrio thermocellus A3DJ38
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.1.78 Ni-agarose column chromatography, and gel filtration Acetivibrio thermocellus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.1.78 ADP + 5'-HO-AUCACGCUUpCp
-
 Acetivibrio thermocellus ?
-
 r
2.7.1.78 ATP + 5'-dephospho-DNA
-
 Acetivibrio thermocellus ADP + 5'-phospho-DNA
-
 r
2.7.1.78 ATP + 5'-dephospho-RNA
-
 Acetivibrio thermocellus ADP + 5'-phospho-RNA
-
 r
2.7.1.78 ATP + 5'-HO-CCGACCAACGAAGGT
-
 Acetivibrio thermocellus ?
-
 r
2.7.1.78 CTP + 5'-dephospho-RNA
-
 Acetivibrio thermocellus CDP + 5'-phospho-RNA
-
 r
2.7.1.78 dATP + 5'-dephospho-RNA
-
 Acetivibrio thermocellus dADP + 5'-phospho-RNA
-
 r
2.7.1.78 GTP + 5'-dephospho-RNA
-
 Acetivibrio thermocellus GDP + 5'-phospho-RNA
-
 r
2.7.1.78 UTP + 5'-dephospho-RNA
-
 Acetivibrio thermocellus UDP + 5'-phospho-RNA
-
 r

Synonyms

EC Number Synonyms Comment Organism
2.7.1.78 PNKP the enzyme is composed of three catalytic modules: an N-terminal 5'-OH polynucleotide kinase, a central 2',3' phosphatase, and a C-terminal ligase polynucleotide kinase Acetivibrio thermocellus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.1.78 0.072
-
 5'-HO-CCGACCAACGAAGGT wild type enzyme, at pH 7.0 and 37°C Acetivibrio thermocellus