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Literature summary for 2.7.1.78 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)

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

Crystallization (Commentary)

Crystallization (Comment) Organism
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

Protein Variants Comment Organism
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]

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

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Acetivibrio thermocellus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + 5'-dephospho-DNA Acetivibrio thermocellus
-
ADP + 5'-phospho-DNA
-
r
ATP + 5'-dephospho-RNA Acetivibrio thermocellus
-
ADP + 5'-phospho-RNA
-
r

Organism

Organism UniProt Comment Textmining
Acetivibrio thermocellus A3DJ38
-
-

Purification (Commentary)

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

Substrates and Products (Substrate)

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

Synonyms

Synonyms Comment Organism
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]

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