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Literature summary extracted from

  • Zhu, H.; Yin, S.; Shuman, S.
    Characterization of polynucleotide kinase/phosphatase enzymes from Mycobacteriophages omega and Cjw1 and vibriophage KVP40 (2004), J. Biol. Chem., 279, 26358-26369.
    View publication on PubMed

Protein Variants

EC Number Protein Variants Comment Organism
2.7.1.78 D169A 50% of wild-type activity with 37-mer oligodeoxyribonucleotide substrate, no reverse reaction Kostyavirus CJW1
2.7.1.78 D175A 60% of wild-type activity with 37-mer oligodeoxyribonucleotide substrate, 68% of wild-type activity with CMP substrate Omegavirus omega
2.7.1.78 K15A no enzymic activity in kinase reaction, 58% of wild-type activity in 3’-phosphates reaction Kostyavirus CJW1
2.7.1.78 K16A no enzymic activity Omegavirus omega
3.1.3.32 D169A half of wild type 5’-kinase activity, almost complete loss of 3’-phosphatase activity Kostyavirus CJW1
3.1.3.32 D175A reduced 5’-kinase activity, no 3’-phosphatase activity Omegavirus omega
3.1.3.32 K15A no 5’-kinase activity, reduced 3’-phosphates activity Kostyavirus CJW1
3.1.3.32 K16A retains 5’-kinase activity, reduced 3’-phosphatase activity Omegavirus omega

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.7.1.78 Mg2+ 1-2 mM, maximum activity, inhibitory above Kostyavirus CJW1
2.7.1.78 Mg2+ 1-2 mM, maximum activity, inhibitory above Omegavirus omega

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.1.78 0.007
-
ATP pH 6.5, 37°C Omegavirus omega
2.7.1.78 0.04
-
ATP pH 6.5, 37°C Kostyavirus CJW1

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.1.78 Co2+ maximum activity at 0.6-2.5 mM Kostyavirus CJW1
2.7.1.78 Mg2+ 1-2 mM, maximum activity, inhibitory above Omegavirus omega
2.7.1.78 Mg2+ about 60% as efficient as Co2+ Kostyavirus CJW1
2.7.1.78 Mn2+ half as effective as Mg2+ Omegavirus omega
2.7.1.78 Mn2+ about 60% as efficient as Co2+ Kostyavirus CJW1
2.7.1.78 additional information divalent metal ion required Omegavirus omega
3.1.3.32 Co2+ divalent cation required, order of decreasing preference: Co2+, Mg2+, Mn2+ Kostyavirus CJW1
3.1.3.32 Co2+ divalent metal ion required, fulfilled by Co2+, Mg2+, Mn2+ at 10 mM Omegavirus omega
3.1.3.32 Mg2+ divalent cation required, order of decreasing preference: Co2+, Mg2+, Mn2+ Kostyavirus CJW1
3.1.3.32 Mg2+ divalent metal ion required, fulfilled by Co2+, Mg2+, Mn2+ at 10 mM Omegavirus omega
3.1.3.32 Mn2+ divalent cation required, order of decreasing preference: Co2+, Mg2+, Mn2+ Kostyavirus CJW1
3.1.3.32 Mn2+ divalent metal ion required, fulfilled by Co2+, Mg2+, Mn2+ at 10 mM Omegavirus omega

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.7.1.78 39000
-
4 * 39000, recombinant protein with His10-tag, calculated, protein is either a stable but asymmetrically shaped homotetramer or a metastable tetramer that is prone to dissociate to a homodimer Kostyavirus CJW1
2.7.1.78 40000
-
4 * 40000, recombinant protein with His10-tag, SDS-PAGE, protein is either a stable but asymmetrically shaped homotetramer or a metastable tetramer that is prone to dissociate to a homodimer Omegavirus omega
2.7.1.78 80000 160000 sedimentation equilibrium analysis Omegavirus omega
2.7.1.78 80000 160000 sedimentaion equilibrium analysis Kostyavirus CJW1
3.1.3.32 40000
-
4 * 40000, His10-tagged enzyme, SDS-PAGE Omegavirus omega
3.1.3.32 80000 160000 sedimentation analysis Omegavirus omega

Organism

EC Number Organism UniProt Comment Textmining
2.7.1.78 Kostyavirus CJW1
-
expression in Escherichia coli with His10-tag
-
2.7.1.78 Omegavirus omega
-
expression in Escherichia coli with His10-tag
-
3.1.3.32 Kostyavirus CJW1
-
bifunctional enzyme with proximal 5’-kinase and disal 3’-phosphatase activity
-
3.1.3.32 Omegavirus omega
-
bifunctional enzyme with proximal 5’-kinase and disal 3’-phosphatase activity
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.1.3.32 19
-
3’-phosphatase activity Omegavirus omega

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.1.78 ATP + 3'-CMP
-
Omegavirus omega ADP + pCp
-
?
2.7.1.78 ATP + 3'-CMP
-
Kostyavirus CJW1 ADP + pCp
-
?
2.7.1.78 ATP + 5'-dephospho-DNA 37-mer oligodeoxyribonucleotide substrate Omegavirus omega ADP + 5'-phospho-DNA
-
r
2.7.1.78 ATP + 5'-dephospho-DNA 37-mer oligodeoxyribonucleotide substrate Kostyavirus CJW1 ADP + 5'-phospho-DNA
-
r
3.1.3.32 dTMP + H2O
-
Omegavirus omega thymidine + phosphate
-
?
3.1.3.32 dTMP + H2O
-
Kostyavirus CJW1 thymidine + phosphate
-
?
3.1.3.32 phosphorylated DNA ends + H2O
-
Omegavirus omega ?
-
?
3.1.3.32 phosphorylated DNA ends + H2O
-
Kostyavirus CJW1 ?
-
?
3.1.3.32 phosphorylated DNA-ends + H2O
-
Omegavirus omega DNA-ends + phosphate
-
r
3.1.3.32 phosphorylated DNA-ends + H2O
-
Kostyavirus CJW1 DNA-ends + phosphate
-
r

Subunits

EC Number Subunits Comment Organism
2.7.1.78 tetramer 4 * 39000, recombinant protein with His10-tag, calculated, protein is either a stable but asymmetrically shaped homotetramer or a metastable tetramer that is prone to dissociate to a homodimer Kostyavirus CJW1
2.7.1.78 tetramer 4 * 40000, recombinant protein with His10-tag, SDS-PAGE, protein is either a stable but asymmetrically shaped homotetramer or a metastable tetramer that is prone to dissociate to a homodimer Omegavirus omega
3.1.3.32 tetramer 4 * 40000, His10-tagged enzyme, SDS-PAGE Omegavirus omega

Synonyms

EC Number Synonyms Comment Organism
3.1.3.32 PNKP
-
Omegavirus omega

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.1.78 0.0043
-
ATP pH 6.5, 37°C Kostyavirus CJW1
2.7.1.78 0.008
-
3'-CMP pH 6.5, 37°C Kostyavirus CJW1
2.7.1.78 0.2
-
ATP pH 6.5, 37°C Omegavirus omega
2.7.1.78 1.6
-
3'-CMP pH 6.5, 37°C Omegavirus omega
3.1.3.32 0.003
-
phosphorylated DNA ends
-
Omegavirus omega
3.1.3.32 0.0043
-
phosphorylated DNA ends
-
Kostyavirus CJW1
3.1.3.32 0.58
-
dTMP
-
Kostyavirus CJW1
3.1.3.32 0.63
-
dTMP
-
Omegavirus omega
3.1.3.32 6.08
-
dTMP
-
Omegavirus omega
3.1.3.32 6.08
-
dTMP
-
Kostyavirus CJW1

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.1.78 5.5 7.5 3’-CMP kinase activit Kostyavirus CJW1
2.7.1.78 8.5 9.5
-
Omegavirus omega
3.1.3.32 5.5
-
-
Omegavirus omega
3.1.3.32 5.5
-
-
Kostyavirus CJW1

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.7.1.78 6
-
50% of maximum activity Omegavirus omega
2.7.1.78 6
-
50% of maximum activity Kostyavirus CJW1